Methods for increasing the stabilization of hypoxia inducible factor-1 alpha

ABSTRACT

Disclosed herein are methods for controlling the activity of hypoxia-inducible transcription factor 1-alpha (HIF-1α) and diseases, conditions, or syndromes related thereto, inter alia, Peripheral Vascular Disease (PVD), Coronary Artery Disease (CAD), heart failure, ischemia, and anemia. Further disclosed are pharmaceutical compositions comprising HIF-1α prolyl hydroxylase inhibitors useful in treating diseases, conditions, and/or syndromes related thereto the activity of HIF-1α.

PRIORITY

This application claims the benefit of Provisional Application Ser. No.61/258,914 and Provisional Application Ser. No. 61/258,918 that wereboth filed on Nov. 6, 2009, the entirety of which applications areincorporated herein by reference.

FIELD OF THE DISCLOSURE

Disclosed herein are prolyl hydroxylase inhibitors that can stabilizehypoxia inducible factor-1 alpha (HIF-1α), as well as hypoxia induciblefactor-2 alpha (HIF-2α). Also disclosed herein are pharmaceuticalcompositions comprising one or more of the disclosed compounds. Yetfurther disclosed are methods for stimulating the cellular immuneresponse in a mammal such as increasing phagocytosis, for example,prolonging the life of phagocytes, inter alia, keratinocytes,neutrophils. As such the disclosed compounds provide methods fortreating diseases that relate to the body's immune response.

SUMMARY

The disclosed compounds stabilize HIF-1α and HIF-2α, as well as otherfactors that are present in a compromised immune system or which aredepleted or over taxed by the presence of a disease state and themanifestations of the disease state, inter alia, sepsis. The disclosedcompounds can be used to treat cancer and can be co-administered withother cancer therapy drugs. In addition, the disclosed compounds can beused to boost the immune response by a mammal when co-administered witha vaccine, for example, flu vaccines, malarial vaccines, yellow fevervaccines, cancer vaccines, and the like.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 depicts the normal metabolic pathway of HIF-1α during normoxia.

FIG. 2 depicts the enhancement of neutrophil killing of S. aureus(Newman strain) with 50 μM and 200 μM of a compound disclosed in TableVIII versus control (DMSO) at 60 and 90 minutes.

FIG. 3 depicts the enhancement of human monocyte cell line (U937)against S. aureus (Newman strain) by 10 μM a compound disclosed in TableVIII versus untreated samples.

FIG. 4 depicts the average percent surviving bacteria in treated vs.untreated U937 cells after infection with S. aureus (Newman strain)after 1 hour pre-treatment (black) or 2 hour (hatched) pre-treatmentwith 10 μM of a compound disclosed in Table VIII.

FIG. 5 depicts the average percent surviving bacteria in treated vs.untreated U937 cells after infection with two strains of S. aureus,Newman (black) or methicillin resistant S. aureus (MRSA) (hatched),after 1 hour pre-treatment with 10 μM of a compound disclosed in TableVIII.

FIG. 6 depicts the average percent surviving bacteria in treated vs.untreated U397 cells after infection with two strains of S. aureus,Newman (black) or MRSA (hatched) and treatment with 10 μM of a compounddisclosed in Table VIII.

FIG. 7 depicts the average percent surviving bacteria in treated vs.untreated U937 cells after infection with two strains of S. aureus,Newman (hatched bars) or MRSA (black bars), following treatment with 100mM mimosine (A), 10 μM of a compound disclosed in Table VIII (B), or 2mg/mL of vancomycin (C) at 2 hours post-infection.

FIG. 8 depicts the average percent surviving bacteria in treated vs.untreated U937 cells after infection with S. aureus (Newman) followingno pre-treatment, 1 hour pre-treatment, or 2 hour pre-treatment with 10μM of a compound disclosed in Table VIII.

FIG. 9 depicts the average percent surviving bacteria in treated vs.untreated HaCaT cells infected with two strains of S. aureus, Newman(hatched bars) or MRSA (black bars) and pre-treated for 1 hour witheither DMSO (control), 800 μM mimosine, 10 μM a compound disclosed inTable VIII or 1 μg/mL vancomycin. Data shown is 2 hours post-treatment.

FIG. 10 depicts the average percent surviving bacteria in treated vs.untreated HaCaT cells infected with two strains of S. aureus, Newman(hatched bars) or MRSA (black bars), following pre-treatment with 10 μMa compound disclosed in Table VIII.

FIG. 11 depicts the up regulation of phosphoglycerate kinase (PGK)expression in wild type murine embryonic fibroblasts as a result oftreatment with a compound disclosed in Table VIII at dosages of 1 μM(E), 10 μM (F), and 50 μM (G) vs. wild type control (H) and the lack ofup regulation of PGK expression in HIF-1 knock out cells as a result oftreatment with a compound disclosed in Table VIII at dosages of 1 μM(A), 10 μM (B), and 50 μM (C) and HIF-1 knock out control (D). Both celltypes were treated for 7 hours.

FIG. 12 depicts the up regulation of phosphoglycerate kinase (PGK)expression in wild type murine embryonic fibroblasts as a result oftreatment with compound I-(3-Chlorobenzyl)-3-hydroxypyridin-2(1H)-one atdosages of 1 μM (E), 10 μM (F), vs. wild type control (G) and the lackof up regulation of PGK expression in HIF-1 knock out cells as a resultof treatment with a compound disclosed in Table VIII at dosages of 1 μM(A), 10 μM (B), and 50 μM (C) and HIF-1 knock out control (D).

FIG. 13 depicts the up regulation of phosphoglycerate kinase (PGK)expression in wild type murine embryonic fibroblasts as a result oftreatment with a compound disclosed in Table VIII at dosages of 1 μM(E), 10 μM (F), and 50 μM (G) vs. wild type control (H) and the lack ofup regulation of PGK expression in HIF-1 knock out cells as a result oftreatment with a compound disclosed in Table VIII at dosages of 1 μM(A), 10 μM (B), and 50 μM (C) and HIF-1 knock out control (D).

FIG. 14 depicts the up regulation of vascular endothelia growth factor(VEGF) expression in wild type murine embryonic fibroblasts as a resultof treatment with a compound disclosed in Table VIII at dosages of 1 μM(E), 10 μM (F), and 50 μM (G) vs. control (H) and the lack of upregulation of VEGF expression in HIF-1 knock out cells treated with acompound disclosed in Table VIII at dosages of 1 μM (A), 10 μM (B), and50 μM (C) and HIF-1 knock out control (D). Both cell types were treatedfor 7 hours.

FIG. 15 depicts the results of Example 11 wherein 3 groups of animalsare treated with Staphylococcus aureus antibiotic sensitive Newmanstrain. The data show the significant reduction in the size of skinlesions (wounds) for animals in Group 1 (solid circles (●)) treated with10 μM of a compound disclosed in Table VIII versus animal given a bolusof DMSO (solid squares (▪)). FIG. 15, depicts mice infected with Newmanstrain of S. aureus followed by treatment with 10 μM of a compounddisclosed in Table VIII or DMSO (control) at 2 hours post-infection. Thedata show the statistically significant reduction in the size of skinlesions (wounds) for animals treated with a disclosed in Table VIII(solid circles (●)) or DMSO (solid squares (▪)).

FIG. 16 also depicts the results of Example 11 showing the reduction inthe size of skin lesions (wounds) for animals in Group 1 (solid circles(●)) treated with 10 μM a compound disclosed in Table VIII versusanimals that are untreated (solid triangles (▴)). FIG. 16 depicts miceinfected with Newman strain of S. aureus followed by treatment with 10μM of a compound disclosed in Table VIII or no treatment at 2 hourspost-infection. The data show the reduction in the size of skin lesions(wounds) for animals treated with a compound disclosed in Table VIII(solid circles (●)) or untreated (solid triangles (▴)).

FIG. 17 is a plot histogram that depicts the results of Example 12wherein 3 groups of animals are treated with Staphylococcus aureusantibiotic sensitive Newman strain [ATCC #25904]. The data show theresults for the untreated group plotted under (A), the results for thegroup treated with DMSO plotted under (B) and results for the grouptreated with 10 μM of a compound disclosed in Table VIII plotted under(C).

FIG. 18 also depicts the results of Example 12 wherein the number ofcolony forming units in the kidney are plotted for the various groups:the untreated group is plotted under (A), the group treated with DMSO isplotted under (B) and the group treated with 10 μM of a compounddisclosed in Table VIII is plotted under (C).

FIG. 19 depicts the results of Example 13 wherein 2 groups of animalsare treated with Streptococcus pyogenes NZ131 [M49 strain]. The datashow the reduction in the size of skin lesions (wounds) for animals inGroup 1 (solid triangles (▴)) treated with 0.5 mg/kg of a compounddisclosed in Table VIII versus animal treated with vehicle control(cyclodextran) (solid circles (●)).

FIG. 20 is a plot histogram that also depicts the results of Example 12wherein the number of colony forming units for the observed skin lesionson animals treated with vehicle control (cyclodextran) are plotted under(A) and results for the group treated with 0.5 mg/kg of a compounddisclosed in Table VIII are plotted under (B).

DETAILED DISCLOSURE

In this specification and in the claims that follow, reference will bemade to a number of terms that shall be defined to have the followingmeanings:

Throughout this specification, unless the context requires otherwise,the word “comprise,” or variations such as “comprises” or “comprising,”will be understood to imply the inclusion of a stated integer or step orgroup of integers or steps but not the exclusion of any other integer orstep or group of integers or steps.

It must be noted that, as used in the specification and the appendedclaims, the singular forms “a,” “an” and “the” include plural referentsunless the context clearly dictates otherwise. Thus, for example,reference to “a carrier” includes mixtures of two or more such carriers,and the like.

“Optional” or “optionally” means that the subsequently described eventor circumstance can or cannot occur, and that the description includesinstances where the event or circumstance occurs and instances where itdoes not.

By “pharmaceutically acceptable” is meant a material that is notbiologically or otherwise undesirable, i.e., the material can beadministered to an individual along with the relevant active compoundwithout causing clinically unacceptable biological effects orinteracting in a deleterious manner with any of the other components ofthe pharmaceutical composition in which it is contained. Ranges may beexpressed herein as from “about” one particular value, and/or to “about”another particular value. When such a range is expressed, another aspectincludes from the one particular value and/or to the other particularvalue. Similarly, when values are expressed as approximations, by use ofthe antecedent “about,” it will be understood that the particular valueforms another aspect. It will be further understood that the endpointsof each of the ranges are significant both in relation to the otherendpoint, and independently of the other endpoint.

A weight percent of a component, unless specifically stated to thecontrary, is based on the total weight of the formulation or compositionin which the component is included.

By “effective amount” as used herein means “an amount of one or more ofthe disclosed HIF-1α prolyl hydroxylase inhibitors, effective at dosagesand for periods of time necessary to achieve the desired or therapeuticresult.” An effective amount may vary according to factors known in theart, such as the disease state, age, sex, and weight of the human oranimal being treated. Although particular dosage regimes may bedescribed in examples herein, a person skilled in the art wouldappreciated that the dosage regime may be altered to provide optimumtherapeutic response. For example, several divided doses may beadministered daily or the dose may be proportionally reduced asindicated by the exigencies of the therapeutic situation. In addition,the compositions of this disclosure can be administered as frequently asnecessary to achieve a therapeutic amount.

“Admixture” or “blend” is generally used herein means a physicalcombination of two or more different components

“Excipient” is used herein to include any other compound that may becontained in or combined with one or more of the disclosed inhibitorsthat is not a therapeutically or biologically active compound. As such,an excipient should be pharmaceutically or biologically acceptable orrelevant (for example, an excipient should generally be non-toxic to thesubject). “Excipient” includes a single such compound and is alsointended to include a plurality of excipients.

As used herein, by a “subject” is meant an individual. Thus, the“subject” can include domesticated animals (e.g., cats, dogs, etc.),livestock (e.g., cattle, horses, pigs, sheep, goats, etc.), laboratoryanimals (e.g., mouse, rabbit, rat, guinea pig, etc.), and birds.“Subject” can also include a mammal, such as a primate or a human.

By “prevent” or other forms of the word, such as “preventing” or“prevention,” is meant to stop a particular event or characteristic, tostabilize or delay the development or progression of a particular eventor characteristic, or to minimize the chances that a particular event orcharacteristic will occur. Prevent does not require comparison to acontrol as it is typically more absolute than, for example, reduce. Asused herein, something could be reduced but not prevented, but somethingthat is reduced could also be prevented. Likewise, something could beprevented but not reduced, but something that is prevented could also bereduced. It is understood that where reduce or prevent are used, unlessspecifically indicated otherwise, the use of the other word is alsoexpressly disclosed.

By “reduce” or other forms of the word, such as “reducing” or“reduction,” is meant lowering of an event or characteristic (e.g.,vascular leakage). It is understood that this is typically in relationto some standard or expected value, in other words it is relative, butthat it is not always necessary for the standard or relative value to bereferred to.

The term “Treat” or other forms of the word such as “treated” or“treatment” is used herein to mean that administration of a compound ofthe present invention mitigates a disease or a disorder in a host and/orreduces, inhibits, or eliminates a particular characteristic or eventassociated with a disorder (e.g., infection caused by a microorganism).Thus, the term “treatment” includes, preventing a disorder fromoccurring in a host, particularly when the host is predisposed toacquiring the disease, but has not yet been diagnosed with the disease;inhibiting the disorder; and/or alleviating or reversing the disorder.Insofar as the methods of the present invention are directed topreventing disorders, it is understood that the term “prevent” does notrequire that the disease state be completely thwarted. Rather, as usedherein, the term preventing refers to the ability of the skilled artisanto identify a population that is susceptible to disorders, such thatadministration of the compounds of the present invention may occur priorto onset of a disease. The term does not imply that the disease state becompletely avoided.

Ranges can be expressed herein as from “about” one particular value,and/or to “about” another particular value. When such a range isexpressed, another aspect includes from the one particular value and/orto the other particular value. Similarly, when values are expressed asapproximations, by use of the antecedent “about,” it will be understoodthat the particular value forms another aspect. It will be furtherunderstood that the endpoints of each of the ranges are significant bothin relation to the other endpoint, and independently of the otherendpoint. It is also understood that there are a number of valuesdisclosed herein, and that each value is also herein disclosed as“about” that particular value in addition to the value itself. Forexample, if the value “10” is disclosed, then “about 10” is alsodisclosed. It is also understood that when a value is disclosed, then“less than or equal to” the value, “greater than or equal to the value,”and possible ranges between values are also disclosed, as appropriatelyunderstood by the skilled artisan. For example, if the value “10” isdisclosed, then “less than or equal to 10” as well as “greater than orequal to 10” is also disclosed. It is also understood that throughoutthe application data are provided in a number of different formats andthat this data represent endpoints and starting points and ranges forany combination of the data points. For example, if a particular datapoint “10” and a particular data point “15” are disclosed, it isunderstood that greater than, greater than or equal to, less than, lessthan or equal to, and equal to 10 and 15 are considered disclosed aswell as between 10 and 15. It is also understood that each unit betweentwo particular units are also disclosed. For example, if 10 and 15 aredisclosed, then 11, 12, 13, and 14 are also disclosed. By“antimicrobial” is meant the ability to treat or control (e.g., reduce,prevent, inhibit, break-down, or eliminate) microorganism growth orsurvival at any concentration. Similarly, the terms “antibacterial,”“antiviral,” and “antifungal” respectively mean the ability to treat orcontrol (e.g., reduce, prevent, inhibit, break-down, or eliminate)bacterial, viral, and fungal growth or survival at any concentration.

The term “anion” is a type of ion and is included within the meaning ofthe term “ion”. An “anion” is any molecule, portion of a molecule (e.g.,zwitterion), cluster of molecules, molecular complex, moiety, or atomthat contains a net negative charge or that can be made to contain a netnegative charge. The term “anion precursor” is used herein tospecifically refer to a molecule that can be converted to an anion via achemical reaction (e.g., deprotonation).

The term “cation” is a type of ion and is included within the meaning ofthe term “ion”. A “cation” is any molecule, portion of a molecule (e.g.,zwitterion), cluster of molecules, molecular complex, moiety, or atom,that contains a net positive charge or that can be made to contain a netpositive charge. The term “cation precursor” is used herein tospecifically refer to a molecule that can be converted to a cation via achemical reaction (e.g., protonation or alkylation).

“Chemotherapeutic agent” is used herein to include any otherpharmaceutically active compound that can be used in conjunction withthe disclosed HIF-1α prolyl hydroxylase inhibitors, for example,cytotoxic drugs such as 6-hydroxymethylacylfulvene, cyclophosphamide,dacarbazine, carmustine, doxorubicin, and methotrexate. Otherchemotherapeutic agents also include anti-inflammatory drugs, i.e.,non-steroidal anti-inflammatory compounds such as aspirin.

Unless stated to the contrary, a formula with chemical bonds shown onlyas solid lines and not as wedges or dashed lines contemplates eachpossible isomer, e.g., each enantiomer, diastereomer, and meso compound,and a mixture of isomers, such as a racemic or scalemic mixture.

The transcription factor Hypoxia-Inducible Factor 1 (HIF-1) is one ofthe key regulators of oxygen homeostasis. It regulates the physiologicalresponses to low oxygen levels (hypoxia) and the pathophysiology ofheart attack, cancer, stroke and chronic lung disease. HIF-1 is aheterodimeric protein that consists of two subunits, HIF-1α and HIF-1β.Whereas HIF-1β is constitutively expressed, the expression of HIF-1α isinduced by oxygen concentrations below 6%. HIF-1 heterodimers bind tothe hypoxia response element (HRE), a 5-RCGTG-3 consensus sequence.Several dozen HIF-1-regulated genes have been identified so far,including genes coding for proteins involved in angiogenesis, energymetabolism, erythropoiesis, cell proliferation and viability, vascularremodeling and vasomotor responses. Therefore, modulation of HIFactivation in cells is critical to preventing, controlling, curing, orotherwise affecting a wide array of diseases, disease states, andconditions.

Hypoxia-inducible transcription factor 1-alpha (HIF-1α) plays a centralrole in cellular adaptation to reduced oxygen availability. Underhypoxic stress, activated HIF-1α strives for oxygen homeostasis by notonly maintaining intracellular energy production via the induction ofangiogenesis and glycolysis, but also limiting energy consumption byvirtue of the inhibition of cell proliferation and DNA repair. Ingeneral, HIF-1α activates its target genes, inter alia, EPO, VEGF, andPGK1 through binding to the hypoxia-responsive element in the genepromoter (Wang, G. L. et al., J Biol Chem (1993); 268: 21513-21518).

HIF-1α under normal healthy conditions wherein the cells have asufficient supply of oxygen is readily converted to a degraded form byone of several 4-prolyl hydroxylase enzymes, inter alia, EGLN1 (hereinreferred to as HIFPH2). As stated above, when cells undergo hypoxia,this enzymatic transformation is slow or entirely stopped andHIF-1αbegins to build up in the cell. When this build up of HIF-1αoccurs, this protein combines with HIF-1β to form the activetranscription factor complex HIF-1. This transcription factor thenactivates several biological pathways which are present as a response toand a means for alleviating the body's state of hypoxia. These responsesinclude, inter alia, angiogenic, erythropoietic (EPO), glucosemetabolism (PGK), matrix alteration, and enhanced capacity of phagocytesto respond to pathogens.

FIG. 1 summaries the metabolism of HIF-1α during normal healthyconditions. The HIF α-subunits are unstable under normoxic conditions;cells continually synthesize and degrade these proteins. The shorthalf-life of HIF-1α is the byproduct of a family of O₂- andiron-dependent prolyl hydroxylases (PH1-3), whose action directs HIFα-subunits for degradation by the ubiquitin-proteasome pathway in aprocess dependent upon interaction with von Hippel-Lindautumor-suppressor protein (vHL). In FIG. 1, PHD's represents the prolylhydroxylases that act in the presence of an asparaginyl hydroxylase tohydroxylate prolines 402 and 564, as well as asparagines 804. From thispoint, because the hydroxylated HIF-1α is also prevented fromassociation with p300-CPB because of other factors, ubiquitin ligasebegins to metabolize the hydroxylated HIF-1α via the vHL pathway.

In patients where there is a need for stimulating this response, forexample, in patients in need of increased tissue oxygen due toperipheral vascular disease (PVD), inhibiting the HIF1 enzymes, forexample, Eg1 nine homolog 1 (HIFPH2), will stimulate the body's ownangiogenic response without the consequences of oxygen deficiency. Inaddition, in diseases of ischemia, inter alia, CAD and anemia,stimulation of angiogenic, erythropoietic, and metabolism adaption canprovide therapeutic benefits. Up regulation of HIF-1α also provides amethod for enhancing immunity, for example, by increasing the capacityof phagocytes.

There is therefore a long felt need for methods for controlling theactivity of HIF-1αwhich can be effectively accomplished by compoundsthat inhibit the 4-prolyl hydroxylase enzymes that degrade HIF-1α. Thisinhibition of 4-prolyl hydroxylase enzymes, inter alia, HIFPH2 (alsoreferred to herein as EGLN1 or PHD2) and HIFPH3 (also referred to hereinas EGLN3 of PHD-3) thereby provide a method for increasing theconcentration of HIF-1α in cells and thus providing methods for treatinga variety of diseases or disease states.

Disclosed herein are methods for treating one or more diseases,conditions, syndromes, and the like that are affected by the level ofhypoxia-inducible transcription factors. Regulation of these factorsboth during hypoxia and normoxia can provide methods for re-balancing orregulating one or more biological pathways associated with abnormalconditions, inter alia, invasion of the body by pathogens, inter alia,bacteria, fungi, viruses, and parasites, abnormal cellular regulation,i.e., cancer ischemia, and the side effects caused by vaccination.

Targeting HIF1 Stabilization in Cells

HIF-1α is targeted for destruction via prolyl hydroxylation, anoxygen-dependent modification that signals for recognition by the E3ubiquitin ligase complex containing the von Hippel-Lindau tumorsuppressor (VHL). Three prolyl hydroxylases formerly referred to in theliterature as EGLN1, EGLN2, and EGLN3 (also known as, have beenidentified in mammals, among which, EGLN1 (also known as HIFPH2 orPHD2), and EGLN3 (also known as HIFPH3 or PHD3), are hypoxia-inducibleat their mRNA levels in a HIF-1α dependent manner. HIF-1α levels arecontrolled by these prolyl-4-hydroxylases by hydroxylating the HIF-1αproline residues Pro-402 and Pro-564 in humans (Ivan, M. et al., (2001)“HIFα targeted for VHL-mediated destruction by proline hydroxylation:implications for O₂ sensing.” Science 292, 464-468; Jaakkola, P. et al.,(2001) “Targeting of HIF-1α to the von Hippel-Lindau ubiquitylationcomplex by O₂-regulated prolyl hydroxylation.” Science 292, 468-472; andMasson, N. et al., (2001) “Independent function of two destructiondomains in hypoxia-inducible factor-α chains activated by prolylhydroxylation.” EMBO J. 20, 5197-5206). Under hypoxia conditions, EGLN1and EGLN3 activity is suppressed.

Stimulated by a build up of the cellular concentration of HIF-1α is theproduction of Phosphoglycerate Kinase (PGK) and Vascular EndothelialGrowth Factor (VEGF). It has been shown that stimulation of VEGF inducesthe formation of functional neo-vessels in the mouse cornea and enhancedblood flow in a dog model of coronary artery disease. The HIF-1α prolylhydroxylase inhibitors of the present disclosure provide enhancement inthe expression of multiple hypoxia inducible genes including VEGF, GAPDHand erythropoietin (EPO). Additionally, the HIF-1α prolyl hydroxylaseinhibitors of the present disclosure provide enhanced the accumulationof HIF1-α in the cytoplasm and nucleus. Transgenic mice expressing aconstitutively active HIF-1α in the skin have increased dermalvascularity and had a 13-fold increase in VEGF levels

Wounds

Chronic, non-healing wounds are a major cause of prolonged morbidity inthe aged human population. This is especially the case in bedridden ordiabetic patients who develop severe, non-healing skin ulcers. In manyof these cases, the delay in healing is a result of inadequate bloodsupply either as a result of continuous pressure or of vascularblockage. Poor capillary circulation due to small artery atherosclerosisor venous stasis contributes to the failure to repair damaged tissue.Such tissues are often infected with microorganisms that proliferateunchallenged by the innate defense systems of the body which requirewell vascularized tissue to effectively eliminate pathogenic organisms.As a result, most therapeutic intervention centers on restoring bloodflow to ischemic tissues thereby allowing nutrients and immunologicalfactors access to the site of the wound.

The present disclosure relates to methods for treating wounds andpromoting wound healing in a subject comprising, administering to asubject in need of treatment an effective amount of one or more of thedisclosed compounds.

The present disclosure relates to the use of one or more of thedisclosed compounds for use in making a medicament for treating woundsand promoting wound healing.

Antimicrobial

The hypoxia-responsive transcription factor HIF-1α is essential forregulation of inflammation in vivo. As such, it has been discovered(Peyssonnaux C. et al., “HIF-1αexpression regulates the bactericidalcapacity of phagocytes” J. Clinical Investigation 115(7), pp 1808-1815(2005)) that bacterial infection induces HIF-1α expression in myeloidcells even under normoxic conditions, and that HIF-1α regulates thegeneration of critical molecular effectors of immune defense includinggranule proteases, antimicrobial peptides, nitric oxide, and TNF-α.Bacterial infection induces a subset of HIF-1α target genes specificallyrelated to microbial killing, thereby demonstrating that HIF-1α has anessential function in innate immunity distinct from hypoxic response.Therefore, HIF-1αfunction is critical for myeloid cell bactericidalactivity and the ability of the host to limit systemic spread ofinfection from an initial tissue focus. Increased activity of theHIF-1αpathway through vHL deletion supports myeloid cell production ofdefense factors and improves bactericidal capacity. The disclosedcompounds induce HIF-1α activity and can also boost bacterial killingand NO production in a HIF-1α-specific fashion. These discoveriesprovide methods for enhancing innate immune responses to microbial, forexample, bacterial, infection.

Without wishing to be limited by theory, the disclosed compounds canincrease the stabilization of HIF-1 protein by acting directly orindirectly on one or more cellular processes which act to destabilize orto metabolize cellular components that stabilize the presence of HIF-1protein, protect it from inhibition, or to increase the activity of theprotein. Alternatively, the disclosed compounds can increase theactivity of the HIF-1 protein by inhibiting or otherwise blocking theactivity of compounds that inhibit the activity of the HIF-1 protein. Assuch, disclosed herein is a method for improving the treatment ofmicrobial infections by administering a substance that increases theactivity or level of at least one HIF-1 protein in a subject sufferingfrom the microbial infection or at increased risk of microbialinfection.

In one aspect, disclosed herein are methods for modulating the activityof at least one HIF-1 protein. As such, the disclosed methods comprisecontacting at least one HIF-1 protein or HIF-1 interacting protein withone or more of the disclosed compounds that modulate the activity of theHIF-1 protein, or causing contact between the protein and substance. Inthe embodiment, the contacting is accomplished in vitro. In anotherembodiment, the contacting is accomplished in vivo. In a furtherembodiment, the contacting is accomplished ex vivo.

In another aspect, disclosed herein is a method of treating a subjectinfected or at risk of infection by a microbial agent comprisingadministering to a subject a therapeutically effective amount of one ormore of the disclosed compounds. In one embodiment, the compoundincreases the amount or activity of HIF-1. In another embodiment, themicrobial agent is a pathogen. Iterations of this embodiment related topathogens includes, bacteria, fungi, protozoa, viruses, yeasts, and thelike. A yet further iteration of this aspect relates to a method fortreating a subject infected by or at risk of infection by a microbialagent comprising, increasing the microbial pathogen-killing activity ofthe subject's immune cells.

One method for increasing the stabilization of HIF-1 is to inhibit theactivity of 4-prolyl hydroxylase enzymes that begin the cellular breakdown of HIF-1α thereby preventing HIF-1α from combining with HIF-1β toform HIF-1. As such, disclosed herein are methods for increasing thecellular response to disease states such as infection, i.e., presence ofa pathogen such as a bacterium, a virus, a parasite, a yeast, a fungus,and the like by increasing phagocytosis. Also disclosed herein aremethods for treating cancer by increasing the cellular immune response,for example, by stabilizing HIF-1, thereby increasing the ability of thebody to reduce tumor size. Further disclosed herein are methods fortreating diseases wherein an immune response can be stimulated byvaccination.

The following chemical hierarchy is used throughout the specification todescribe and enable the scope of the present disclosure and toparticularly point out and distinctly claim the units which comprise thecompounds of the present disclosure, however, unless otherwisespecifically defined, the terms used herein are the same as those of theartisan of ordinary skill. The term “hydrocarbyl” stands for any carbonatom-based unit (organic molecule), said units optionally containing oneor more organic functional group, including inorganic atom comprisingsalts, inter alia, carboxylate salts, quaternary ammonium salts. Withinthe broad meaning of the term “hydrocarbyl” are the classes “acyclichydrocarbyl” and “cyclic hydrocarbyl” which terms are used to dividehydrocarbyl units into cyclic and non-cyclic classes.

As it relates to the following definitions, “cyclic hydrocarbyl” unitscan comprise only carbon atoms in the ring (carbocyclic and aryl rings)or can comprise one or more heteroatoms in the ring (heterocyclic andheteroaryl). For “carbocyclic” rings the lowest number of carbon atomsin a ring are 3 carbon atoms; cyclopropyl. For “aryl” rings the lowestnumber of carbon atoms in a ring are 6 carbon atoms; phenyl. For“heterocyclic” rings the lowest number of carbon atoms in a ring is 1carbon atom; diazirinyl. Ethylene oxide comprises 2 carbon atoms and isa C₂ heterocycle. For “heteroaryl” rings the lowest number of carbonatoms in a ring is 1 carbon atom; 1,2,3,4-tetrazolyl. The following is anon-limiting description of the terms “acyclic hydrocarbyl” and “cyclichydrocarbyl” as used herein.

A. Substituted and Unsubstituted Acyclic Hydrocarbyl:

-   -   For the purposes of the present disclosure the term “substituted        and unsubstituted acyclic hydrocarbyl” encompasses 3 categories        of units:

-   1) linear or branched alkyl, non-limiting examples of which include,    methyl (C₁), ethyl (C₂), n-propyl (C₃), iso-propyl (C₃), n-butyl    (C₄), sec-butyl (C₄), iso-butyl (C₄), tert-butyl (C₄), and the like;    substituted linear or branched alkyl, non-limiting examples of which    includes, hydroxymethyl (C₁), chloromethyl (C₁), trifluoromethyl    (C₁), aminomethyl (C₁), 1-chloroethyl (C₂), 2-hydroxyethyl (C₂),    1,2-difluoroethyl (C₂), 3-carboxypropyl (C₃), and the like.

-   2) linear or branched alkenyl, non-limiting examples of which    include, ethenyl (C₂), 3-propenyl (C₃), 1-propenyl (also    2-methylethenyl) (C₃), isopropenyl (also 2-methylethen-2-yl) (C₃),    buten-4-yl (C₄), and the like; substituted linear or branched    alkenyl, non-limiting examples of which include, 2-chloroethenyl    (also 2-chlorovinyl) (C₂), 4-hydroxybuten-1-yl (C₄),    7-hydroxy-7-methyloct-4-en-2-yl (C₉),    7-hydroxy-7-methyloct-3,5-dien-2-yl (C₉), and the like.

-   3) linear or branched alkynyl, non-limiting examples of which    include, ethynyl (C₂), prop-2-ynyl (also propargyl) (C₃),    propyn-1-yl (C₃), and 2-methyl-hex-4-yn-1-yl (C₇); substituted    linear or branched alkynyl, non-limiting examples of which include,    5-hydroxy-5-methylhex-3-ynyl (C₇), 6-hydroxy-6-methylhept-3-yn-2-yl    (C₈), 5-hydroxy-5-ethylhept-3-ynyl (C₉), and the like.    B. Substituted and Unsubstituted Cyclic Hydrocarbyl:    -   For the purposes of the present disclosure the term “substituted        and unsubstituted cyclic hydrocarbyl” encompasses 5 categories        of units:

-   1) The term “carbocyclic” is defined herein as “encompassing rings    comprising from 3 to 20 carbon atoms, wherein the atoms which    comprise said rings are limited to carbon atoms, and further each    ring can be independently substituted with one or more moieties    capable of replacing one or more hydrogen atoms.” The following are    non-limiting examples of “substituted and unsubstituted carbocyclic    rings” which encompass the following categories of units:    -   i) carbocyclic rings having a single substituted or        unsubstituted hydrocarbon ring, non-limiting examples of which        include, cyclopropyl (C₃), 2-methyl-cyclopropyl (C₃),        cyclopropenyl (C₃), cyclobutyl (C₄), 2, 3-dihydroxycyclobutyl        (C₄), cyclobutenyl (C₄), cyclopentyl (C₅), cyclopentenyl (C₅),        cyclopentadienyl (C₅), cyclohexyl (C₆), cyclohexenyl (C₆),        cycloheptyl (C₇), cyclooctanyl (C₈), 2,5-dimethylcyclopentyl        (C₅), 3,5-dichlorocyclohexyl (C₆), 4-hydroxycyclohexyl (C₆), and        3,3,5-trimethylcyclohex-1-yl (C₆).    -   ii) carbocyclic rings having two or more substituted or        unsubstituted fused hydrocarbon rings, non-limiting examples of        which include, octahydropentalenyl (C₈), octahydro-1H-indenyl        (C₉), 3a,4,5,6,7,7a-hexahydro-3H-inden-4-yl (C₉), decalinyl        (C₁₀), decahydroazulenyl (C₁₀).    -   iii) carbocyclic rings which are substituted or unsubstituted        bicyclic hydrocarbon rings, non-limiting examples of which        include, bicyclo-[2.1.1]hexanyl, bicyclo[2.2.1]heptanyl,        bicyclo[3.1.1]heptanyl, 1,3-dimethyl[2.2.1]heptan-2-yl,        bicyclo[2.2.2]octanyl, and bicyclo[3.3.3]undecanyl.

-   2) The term “aryl” is defined herein as “units encompassing at least    one phenyl or naphthyl ring and wherein there are no heteroaryl or    heterocyclic rings fused to the phenyl or naphthyl ring and further    each ring can be independently substituted with one or more moieties    capable of replacing one or more hydrogen atoms.” The following are    non-limiting examples of “substituted and unsubstituted aryl rings”    which encompass the following categories of units:    -   i) C₆ or C₁₀ substituted or unsubstituted aryl rings; phenyl and        naphthyl rings whether substituted or unsubstituted,        non-limiting examples of which include, phenyl (C₆),        naphthylen-1-yl (C₁₀), naphthylen-2-yl (C₁₀), 4-fluorophenyl        (C₆), 2-hydroxyphenyl (C₆), 3-methylphenyl (C₆),        2-amino-4-fluorophenyl (C₆), 2-(N,N-diethylamino)phenyl (C₆),        2-cyanophenyl (C₆), 2,6-di-tert-butylphenyl (C₆),        3-methoxyphenyl (C₆), 8-hydroxynaphthylen-2-yl (C₁₀),        4,5-dimethoxynaphthylen-1-yl (C₁₀), and 6-cyano-naphthylen-1-yl        (C₁₀).    -   ii) C₆ or C₁₀ aryl rings fused with 1 or 2 saturated rings        non-limiting examples of which include,        bicyclo[4.2.0]octa-1,3,5-trienyl (C₈), and indanyl (C₉).

-   3) The terms “heterocyclic” and/or “heterocycle” are defined herein    as “units comprising one or more rings having from 3 to 20 atoms    wherein at least one atom in at least one ring is a heteroatom    chosen from nitrogen (N), oxygen (O), or sulfur (S), or mixtures of    N, O, and S, and wherein further the ring which comprises the    heteroatom is also not an aromatic ring.” The following are    non-limiting examples of “substituted and unsubstituted heterocyclic    rings” which encompass the following categories of units:    -   i) heterocyclic units having a single ring containing one or        more heteroatoms, non-limiting examples of which include,        diazirinyl (C₁), aziridinyl (C₂), urazolyl (C₂), azetidinyl        (C₃), pyrazolidinyl (C₃), imidazolidinyl (C₃), oxazolidinyl        (C₃), isoxazolinyl (C₃), thiazolidinyl (C₃), isothiazolinyl        (C₃), oxathiazolidinonyl (C₃), oxazolidinonyl (C₃), hydantoinyl        (C₃), tetrahydrofuranyl (C₄), pyrrolidinyl (C₄), morpholinyl        (C₄), piperazinyl (C₄), piperidinyl (C₄), dihydropyranyl (C₅),        tetrahydropyranyl (C₅), piperidin-2-onyl (valerolactam) (C₅),        2,3,4,5-tetrahydro-1H-azepinyl (C₆), 2,3-dihydro-1H-indole (C₈),        and 1,2,3,4-tetrahydro-quinoline (C₉).    -   ii) heterocyclic units having 2 or more rings one of which is a        heterocyclic ring, non-limiting examples of which include        hexahydro-1H-pyrrolizinyl (C₇),        3a,4,5,6,7,7a-hexahydro-1H-benzo[d]imidazolyl (C₇),        3a,4,5,6,7,7a-hexahydro-1H-indolyl (C₈),        1,2,3,4-tetrahydroquinolinyl (C₉), and        decahydro-1H-cycloocta[b]pyrrolyl (C₁₀).

-   4) The term “heteroaryl” is defined herein as “encompassing one or    more rings comprising from 5 to 20 atoms wherein at least one atom    in at least one ring is a heteroatom chosen from nitrogen (N),    oxygen (O), or sulfur (S), or mixtures of N, O, and S, and wherein    further at least one of the rings which comprises a heteroatom is an    aromatic ring.” The following are non-limiting examples of    “substituted and unsubstituted heterocyclic rings” which encompass    the following categories of units:    -   i) heteroaryl rings containing a single ring, non-limiting        examples of which include, 1,2,3,4-tetrazolyl (C₁),        [1,2,3]triazolyl (C₂), [1,2,4]triazolyl (C₂), triazinyl (C₃),        thiazolyl (C₃), 1H-imidazolyl (C₃), oxazolyl (C₃), isoxazolyl        (C₃), isothiazolyl (C₃), furanyl (C₄), thiopheneyl (C₄),        pyrimidinyl (C₄), 2-phenylpyrimidinyl (C₄), pyridinyl (C₅),        3-methylpyridinyl (C₅), and 4-dimethylaminopyridinyl (C₅).    -   ii) heteroaryl rings containing 2 or more fused rings one of        which is a heteroaryl ring, non-limiting examples of which        include: 7H-purinyl (C₅), 9H-purinyl (C₅), 6-amino-9H-purinyl        (C₅), 5H-pyrrolo[3,2-d]pyrimidinyl (C₆),        7H-pyrrolo[2,3-d]pyrimidinyl (C₆), pyrido[2,3-d]pyrimidinyl        (C₂), 2-phenylbenzo[d]thiazolyl (C₂), 1H-indolyl (C₈),        4,5,6,7-tetrahydro-1-H-indolyl (C₈), quinoxalinyl (C₈),        5-methylquinoxalinyl (C₈), quinazolinyl (C₈), quinolinyl (C₉),        8-hydroxy-quinolinyl (C₉), and isoquinolinyl (C₉).

-   5) C₁-C₆ tethered cyclic hydrocarbyl units (whether carbocyclic    units, C₆ or C₁₀ aryl units, heterocyclic units, or heteroaryl    units) which connected to another moiety, unit, or core of the    molecule by way of a C₁-C₆ alkylene unit. Non-limiting examples of    tethered cyclic hydrocarbyl units include benzyl C₁-(C₆) having the    formula:

-   -   wherein R^(a) is optionally one or more independently chosen        substitutions for hydrogen. Further examples include other aryl        units, inter alia, (2-hydroxyphenyl)hexyl C₆-(C₆);        naphthalen-2-ylmethyl C₁-(C₁₀), 4-fluorobenzyl C₁-(C₆),        2-(3-hydroxy-phenyl)ethyl C₂-(C₆), as well as substituted and        unsubstituted C₃-C₁₀ alkylenecarbocyclic units, for example,        cyclopropylmethyl C₁-(C₃), cyclopentylethyl C₂-(C₅),        cyclohexylmethyl C₁-(C₆). Included within this category are        substituted and unsubstituted C₁-C₁₀ alkylene-heteroaryl units,        for example a 2-picolyl C₁-(C₆) unit having the formula:

-   -   wherein R^(a) is the same as defined above. In addition, C₁-C₁₂        tethered cyclic hydrocarbyl units include C₁-C₁₀        alkyleneheterocyclic units and alkylene-heteroaryl units,        non-limiting examples of which include, aziridinylmethyl C₁-(C₂)        and oxazol-2-ylmethyl C₁-(C₃).

For the purposes of the present disclosure carbocyclic rings are from C₃to C₂₀; aryl rings are C₆ or C₁₀; heterocyclic rings are from C₁ to C₉;and heteroaryl rings are from C₁ to C₉.

For the purposes of the present disclosure, and to provide consistencyin defining the present disclosure, fused ring units, as well asspirocyclic rings, bicyclic rings and the like, which comprise a singleheteroatom will be characterized and referred to herein as beingencompassed by the cyclic family corresponding to the heteroatomcontaining ring, although the artisan can have alternativecharacterizations. For example, 1,2,3,4-tetrahydroquinoline having theformula:

is, for the purposes of the present disclosure, considered aheterocyclic unit. 6,7-Dihydro-5H-cyclopentapyrimidine having theformula:

is, for the purposes of the present disclosure, considered a heteroarylunit. When a fused ring unit contains heteroatoms in both a saturatedring (heterocyclic ring) and an aryl ring (heteroaryl ring), the arylring will predominate and determine the type of category to which thering is assigned herein for the purposes of describing the disclosure.For example, 1,2,3,4-tetrahydro-[1,8]naphthyridine having the formula:

is, for the purposes of the present disclosure, considered a heteroarylunit.

The term “substituted” is used throughout the specification. The term“substituted” is applied to the units described herein as “substitutedunit or moiety is a hydrocarbyl unit or moiety, whether acyclic orcyclic, which has one or more hydrogen atoms replaced by a substituentor several substituents as defined herein below.” The units, whensubstituting for hydrogen atoms are capable of replacing one hydrogenatom, two hydrogen atoms, or three hydrogen atoms of a hydrocarbylmoiety at a time. In addition, these substituents can replace twohydrogen atoms on two adjacent carbons to form said substituent, newmoiety, or unit. For example, a substituted unit that requires a singlehydrogen atom replacement includes halogen, hydroxyl, and the like. Atwo hydrogen atom replacement includes carbonyl, oximino, and the like.A two hydrogen atom replacement from adjacent carbon atoms includesepoxy, and the like. Three hydrogen replacement includes cyano, and thelike. The term substituted is used throughout the present specificationto indicate that a hydrocarbyl moiety, inter alia, aromatic ring, alkylchain; can have one or more of the hydrogen atoms replaced by asubstituent. When a moiety is described as “substituted” any number ofthe hydrogen atoms can be replaced. For example, 4-hydroxyphenyl is a“substituted aromatic carbocyclic ring (aryl ring)”,(N,N-dimethyl-5-amino)octanyl is a “substituted C₈ linear alkyl unit,3-guanidinopropyl is a “substituted C₃ linear alkyl unit,” and2-carboxypyridinyl is a “substituted heteroaryl unit.”

The following are non-limiting examples of units which can substitutefor hydrogen atoms on a carbocyclic, aryl, heterocyclic, or heteroarylunit:

-   i) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched, or    C₃-C₁₂ cyclic alkyl; for example, methyl (C₁), chloromethyl (C₁),    trifluoromethyl (C₁), aminomethyl (C₁), ethyl (C₂), hydroxymethyl    1-chloroethyl (C₂), 2-hydroxyethyl (C₂), 1,2-difluoroethyl (C₂),    n-propyl (C₃), iso-propyl (C₃), 3-carboxypropyl (C₃), cyclopropyl    (C₃), 2-methyl-cyclopropyl (C₃), n-butyl (C₄), sec-butyl (C₄),    iso-butyl (C₄), tert-butyl (C₄), cyclobutyl (C₄),    2,3-dihydroxycyclobutyl (C₄), pentyl (C₅), cyclopentyl (C₅), hexyl    (C₆), and cyclohexyl (C₆), and the like;-   ii) substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched, or    C₃-C₁₂ cyclic alkenyl; for example, ethenyl (C₂), 2-chloroethenyl    (also 2-chlorovinyl) (C₂), 3-propenyl (C₃), 1-propenyl (also    2-methylethenyl) (C₃), isopropenyl (also 2-methylethen-2-yl) (C₃),    buten-4-yl (C₄), 4-hydroxybuten-1-yl (C₄), cyclobutenyl (C₄),    cyclopentenyl (C₅), cyclopentadienyl (C₅), cyclohexenyl (C₆),    7-hydroxy-7-methyloct-4-en-2-yl (C₉), and    7-hydroxy-7-methyloct-3,5-dien-2-yl (C₉), and the like;-   iii) substituted or unsubstituted C₂-C₁₂ linear or C₃-C₁₂ branched    alkynyl; for example, ethynyl (C₂), prop-2-ynyl (also propargyl)    (C₃), propyn-1-yl (C₃), 2-methyl-hex-4-yn-1-yl (C₇);    5-hydroxy-5-methylhex-3-ynyl (C₇), 6-hydroxy-6-methylhept-3-yn-2-yl    (C₈), 5-hydroxy-5-ethylhept-3-ynyl (C₉), and the like;-   iv) substituted or unsubstituted C₆ or C₁₀ aryl; for example,    phenyl, 2-chlorophenyl, 3-hydroxyphenyl, 4-nitrophenyl,    2-fluoro-4-methylphenyl, 3,5-dinitrophenyl, 8-hydroxynaphth-1-yl,    6-sulfonylnapth-2-yl, and the like;-   v) substituted or unsubstituted C₁-C₉ heterocyclic; for example, as    defined further herein;-   vi) substituted or unsubstituted C₁-C₁₁ heteroaryl; for example, as    defined further herein;-   vii) halogen; for example, fluoro, chloro, bromo, and iodo;-   viii) —[C(R^(23a))(R^(23b))]_(x)OR¹⁰;    -   R¹⁰ is chosen from:    -   a) —H;    -   b) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   c) C₆ or C₁₀ substituted or unsubstituted aryl or alkylenearyl;    -   d) C₁-C₉ substituted or unsubstituted heterocyclic;    -   e) C₁-C₁₁ substituted or unsubstituted heteroaryl;-   ix) —[C(R^(23a))(R^(23b))]_(x)N(R^(11a))(R^(11b));    -   R^(11a) and R^(11b) are each independently chosen from:    -   a) —H;    -   b) —OR¹²;        -   R¹² is hydrogen or C₁-C₄ linear alkyl;    -   c) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   d) C₆ or C₁₀ substituted or unsubstituted aryl;    -   e) C₁-C₉ substituted or unsubstituted heterocyclic;    -   f) C₁-C₁₁ substituted or unsubstituted heteroaryl; or    -   g) R^(11a) and R^(11b) can be taken together to form a        substituted or unsubstituted ring having from 3 to 10 carbon        atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen,        and sulfur;-   x) —[C(R^(23a))(R^(23b))]_(x)C(O)R¹³;    -   R¹³ is:    -   a) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   b) —OR¹⁴;        -   R¹⁴ is hydrogen, substituted or unsubstituted C₁-C₄ linear            alkyl, C₆ or C₁₀ substituted or unsubstituted aryl, C₁-C₉            substituted or unsubstituted heterocyclic, C₁-C₁₁            substituted or unsubstituted heteroaryl;    -   c) —N(R^(15a))(R^(15b));        -   R^(15a) and R^(15b) are each independently hydrogen,            substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,            or C₃-C₁₂ cyclic alkyl; C₆ or C₁₀ substituted or            unsubstituted aryl; C₁-C₉ substituted or unsubstituted            heterocyclic; C₁-C₁₁ substituted or unsubstituted            heteroaryl; or R^(15a) and R^(15b) can be taken together to            form a substituted or unsubstituted ring having from 3 to 10            carbon atoms and from 0 to 3 heteroatoms chosen from oxygen,            nitrogen, and sulfur;-   xi) —[C(R^(23a))(R^(23b))]_(x)OC(O)R¹⁶;    -   R¹⁶ is:    -   a) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   b) —N(R^(17a))(R^(17b));        -   R^(17a) and R^(17b) are each independently hydrogen,            substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,            or C₃-C₁₂ cyclic alkyl; C₆ or C₁₀ substituted or            unsubstituted aryl; C₁-C₉ substituted or unsubstituted            heterocyclic; C₁-C₁₁ substituted or unsubstituted            heteroaryl; or R^(17a) and R^(17b) can be taken together to            form a substituted or unsubstituted ring having from 3 to 10            carbon atoms and from 0 to 3 heteroatoms chosen from oxygen,            nitrogen, and sulfur;-   xii) —[C(R^(23a))(R^(23b))]_(x)NR¹⁸C(O)R¹⁹;    -   R¹⁸ is:    -   a) —H; or    -   b) substituted or unsubstituted C₁-C₄ linear, C₃-C₄ branched, or        C₃-C₄ cyclic alkyl;    -   R¹⁹ is:    -   a) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   b) —N(R^(20a))(R^(20b));        -   R^(20a) and R^(20b) are each independently hydrogen,            substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,            or C₃-C₁₂ cyclic alkyl; C₆ or C₁₀ substituted or            unsubstituted aryl; C₁-C₉ substituted or unsubstituted            heterocyclic; C₁-C₁₁ substituted or unsubstituted            heteroaryl; or R^(20a) and R^(20b) can be taken together to            form a substituted or unsubstituted ring having from 3 to 10            carbon atoms and from 0 to 3 heteroatoms chosen from oxygen,            nitrogen, and sulfur;-   xiii) —[C(R^(23a))(R^(23b))]_(x)CN;-   xiv) —[C(R^(23a))(R^(23b))]_(x)NO₂;-   xv) —[C(R^(23a))(R^(23b))]_(x)R²¹;    -   R²¹ is C₁-C₁₀ linear, C₃-C₁₀ branched, or C₃-C₁₀ cyclic alkyl        substituted by from 1 to 21 halogen atoms chosen from —F, —Cl,        —Br, or —I;-   xvi) —[C(R^(23a))(R^(23b))]_(x)SO₂R²²;    -   R²² is hydrogen, hydroxyl, substituted or unsubstituted C₁-C₄        linear or C₃-C₄ branched alkyl; substituted or unsubstituted C₆,        C₁₀, or C₁₋₄ aryl; C₇-C₁₅ alkylenearyl; C₁-C₉ substituted or        unsubstituted heterocyclic; or C₁-C₁₁ substituted or        unsubstituted heteroaryl;        R^(23a) and R^(23b) are each independently hydrogen or C₁-C₄        alkyl; and        the index x is an integer from 0 to 5.

The compounds disclosed herein include all salt forms, for example,salts of both basic groups, inter alia, amines, as well as salts ofacidic groups, inter alia, carboxylic acids. The following arenon-limiting examples of anions that can form salts with basic groups:chloride, bromide, iodide, sulfate, bisulfate, carbonate, bicarbonate,phosphate, formate, acetate, propionate, butyrate, pyruvate, lactate,oxalate, malonate, maleate, succinate, tartrate, fumarate, citrate, andthe like. The following are non-limiting examples of cations that canform salts of acidic groups: sodium, lithium, potassium, calcium,magnesium, bismuth, and the like.

For the purposes of the present disclosure the terms “compound,”“analog,” and “composition of matter” stand equally well for one anotherand include all enantiomeric forms, diastereomeric forms, salts, and thelike, and the terms “compound,” “analog,” and “composition of matter.”

HIF-1α Prolyl Hydroxylase Inhibitors

The disclosed compounds have the following formulae:

wherein L is chosen from CH₂ or SO₂, thereby providing for N-substitutedbenzyl or N-substituted sulfonylaryl-3-hydroxypyridin-2-(1H)-ones. Y, R¹and R² are further defined herein below.

Disclosed herein are N-substituted benzyl and N-substitutedsulfonylaryl-4-aminomethylene-3-hydroxypyridin-2-(1H)-ones that areHIF-1α prolyl hydroxylase inhibitors having the formula:

wherein R¹ and R² are further defined herein below.

Alkyl piperizine-1-carboxylates

One category of these compounds relates to C₁-C₄ linear or branchedalkyl 4-{[(1-N-(chloro- orfluoro-substituted)-benzyl]-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl)methyl}piperazine-1-carboxylateshaving the formula:

wherein Z is a phenyl group that is substituted with from 1 to 5 halogenatoms that are chosen from chloro and fluoro, and R¹ and R² are takentogether to form a piperazine ring that is substituted with alkylcarboxyunit wherein R⁴ is chosen from C₁-C₄ linear or C₃-C₄ branched alkyl, forexample, ten butyl4{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylatehaving the formula:

One aspect of R⁴ units relates to compounds wherein R⁴ is tert-butyl(C₄). Another aspect of R⁴ units relates to compounds wherein R⁴ ismethyl (C₁). A further aspect of R⁴ units relates to compounds whereinR⁴ is ethyl (C₂). A still further aspect of R⁴ units relates tocompounds wherein R⁴ is chosen from n-propyl (C₃), iso-propyl (C₃),n-butyl (C₄), sec-butyl (C₄), and iso-butyl (C₄). R⁴ is not hydrogen,therefore, a carboxylate unit having the formula: —CO₂H is expresslyexcluded from this category, but may be included in other categories asdescribed herein below.

Z is phenyl substituted with from 1 to 5 halogens chosen from fluorineand chlorine. One aspect of Z units relates to compounds wherein Z is4-chlorophenyl. Another aspect of Z units relates to compounds wherein Zis chosen from 2-chlorophenyl, 3-chlorophenyl, 2-fluorophenyl,3-fluorophenyl, or 4-fluorophenyl. A further aspect of Z units relatesto compounds wherein Z is chosen from 2,3-difluorophenyl,2,4-difluorophenyl, 2,5-difluorophenyl, 2,6-difluorophenyl,2,3-dichlorophenyl, 2,4-dichlorophenyl, 2,5-dichlorophenyl, and2,6-dichlorophenyl.

The following are non-limiting examples of compounds according to thiscategory:

-   methyl    4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   methyl    4-{[1-(3-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   methyl    4-{[1-(2-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   ethyl    4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   ethyl    4-{[1-(3-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   ethyl    4-{[1-(2-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   tert-butyl    4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   tert-butyl    4-{[1-(3-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   tert-butyl    4-{[1-(2-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   methyl    4-{[1-(4-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   methyl    4-{[1-(3-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   methyl    4-{[1-(2-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   ethyl    4-{[1-(4-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   ethyl    4-{[1-(3-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   ethyl    4-{[1-(2-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   tert-butyl    4-{[1-(4-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

-   tert-Butyl    4-{[1-(3-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylate    having the formula:

and

-   tert-butyl    4-{[1-(2-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate    having the formula:

Another category of compounds relates toN-unsubstituted-benzyl-4-aminomethyl-3-hydroxypyridin-2-(1H)-ones,wherein Z is an unsubstituted phenyl group, having the formula:

wherein R¹ and R² are be taken together to form a substituted orunsubstituted heterocyclic or heteroaryl ring.

A first aspect of this category relates to compounds having the formula:

wherein R¹ and R² are be taken together to form a substituted orunsubstituted heterocyclic or heteroaryl ring represented by ring Ahaving from 2 to 20 carbon atoms and from 1 to 7 heteroatoms, and R²⁰⁰represents from 0 to 40 substitutions form hydrogen. The index w is aninteger from 0 to 40. Non-limiting examples of rings include diazirinyl(C₁), 1,2,3,4-tetrazolyl (C₁), aziridinyl (C₂), urazolyl (C₂),[1,2,3]triazolyl (C₂), [1,2,4]triazolyl (C₂), azetidinyl (C₃),pyrazolidinyl (C₃), imidazolidinyl (C₃), oxazolidinyl (C₃), isoxazolinyl(C₃), isoxazolyl (C₃), thiazolidinyl (C₃), isothiazolyl (C₃),isothiazolinyl (C₃), oxathiazolidinonyl (C₃), oxazolidinonyl (C₃),hydantoinyl (C₃), 1H-imidazolyl (C₃), pyrrolidinyl (C₄), morpholinyl(C₄), piperazinyl (C₄), piperidinyl (C₄), piperidin-2-onyl(valerolactam) (C₅), 7H-purinyl (C₅), 9H-purinyl (C₅),6-amino-9H-purinyl (C₅), 2,3,4,5-tetrahydro-1H-azepinyl (C₆),5H-pyrrolo[3,2-d]pyrimidinyl (C₆), 7H-pyrrolo[2,3-d]pyrimidinyl (C₆),and 1,2,3,4-tetrahydroquinoline (C₉).

Each R²⁰⁰ unit is independently chosen from:

-   i) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched, or    C₃-C₁₂ cyclic alkyl; for example, methyl (C₁), (C₁), chloromethyl    (C₁), trifluoromethyl (C₁), aminomethyl (C₁), ethyl (C₂),    hydroxymethyl 1-chloroethyl (C₂), 2-hydroxyethyl (C₂),    1,2-difluoroethyl (C₂), n-propyl (C₃), iso-propyl (C₃),    3-carboxypropyl (C₃), cyclopropyl (C₃), 2-methyl-cyclopropyl (C₃),    n-butyl (C₄), sec-butyl (C₄), iso-butyl (C₄), tert-butyl (C₄),    cyclobutyl (C₄), 2,3-dihydroxycyclobutyl (C₄), pentyl (C₅),    cyclopentyl (C₅), hexyl (C₆), and cyclohexyl (C₆), and the like;-   ii) substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched, or    C₃-C₁₂ cyclic alkenyl; for example, ethenyl (C₂), 2-chloroethenyl    (also 2-chlorovinyl) (C₂), 3-propenyl (C₃), 1-propenyl (also    2-methylethenyl) (C₃), isopropenyl (also 2-methylethen-2-yl) (C₃),    buten-4-yl (C₄), 4-hydroxybuten-1-yl (C₄), cyclobutenyl (C₄),    cyclopentenyl (C₅), cyclopentadienyl (C₅), cyclohexenyl (C₆),    7-hydroxy-7-methyloct-4-en-2-yl (C₉), and    7-hydroxy-7-methyloct-3,5-dien-2-yl (C₉), and the like;-   iii) substituted or unsubstituted C₁-C₁₂ linear or C₃-C₁₂ branched    alkynyl; for example, ethynyl (C₂), prop-2-ynyl (also propargyl)    (C₃), propyn-1-yl (C₃), 2-methyl-hex-4-yn-1-yl (C₇);    5-hydroxy-5-methylhex-3-ynyl (C₇), 6-hydroxy-6-methylhept-3-yn-2-yl    (C₈), 5-hydroxy-5-ethylhept-3-ynyl (C₉), and the like;-   iv) substituted or unsubstituted C₆ or C₁₀ aryl; for example, phenyl    (C₆), naphthylen-1-yl (C₁₀), naphthylen-2-yl (C₁₀), 4-fluorophenyl    (C₆), 2-hydroxyphenyl (C₆), 3-methylphenyl (C₆),    2-amino-4-fluorophenyl (C₆), 2-(N,N-diethylamino)phenyl (C₆),    2-cyanophenyl (C₆), 2,6-di-tert-butylphenyl (C₆), 3-methoxyphenyl    (C₆), 8-hydroxynaphthylen-2-yl (C₁₀), 4,5-dimethoxynaphthylen-1-yl    (C₁₀), 6-cyano-naphthylen-1-yl (C₁₀), and the like;-   v) substituted or unsubstituted C₁-C₉ heterocyclic; for example,    diazirinyl (C₁), aziridinyl (C₂), urazolyl (C₂), azetidinyl (C₃),    pyrazolidinyl (C₃), imidazolidinyl (C₃), oxazolidinyl (C₃),    isoxazolinyl (C₃), isoxazolyl (C₃), thiazolidinyl (C₃), isothiazolyl    (C₃), isothiazolinyl (C₃), oxathiazolidinonyl (C₃), oxazolidinonyl    (C₃), hydantoinyl (C₃), tetrahydropyranyl (C₄), pyrrolidinyl (C₄),    morpholinyl (C₄), piperazinyl (C₄), piperidinyl (C₄), dihydropyranyl    (C₅), tetrahydropyranyl (C₅), piperidin-2-onyl (valerolactam) (C₅),    and the like;-   vi) substituted or unsubstituted C₁-C₁₁ heteroaryl; for example,    1,2,3,4-tetrazolyl (C₁), [1,2,3]triazolyl (C₂), [1,2,4]triazolyl    (C₂), triazinyl (C₃), thiazolyl (C₃), 1H-imidazolyl (C₃), oxazolyl    (C₃), furanyl (C₄), thiopheneyl (C₄), pyrimidinyl (C₄), pyridinyl    (C₅), and the like;-   vii) halogen; for example, —F, —Cl, —Br, or —I;-   viii) —[C(R^(37a))(R^(37b))]_(y)OR²⁴;    -   R²⁴ is chosen from:    -   a) —H;    -   b) substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   c) substituted or unsubstituted C₆ or C₁₀ aryl or C₇ or C₁₀        alkylenearyl; for example, phenyl or benzyl    -   d) substituted or unsubstituted C₁-C₉ heterocyclic;    -   e) substituted or unsubstituted C₁-C₁₁ heteroaryl;        -   for example, —OH, —CH₂OH, —OCH₃, —CH₂OCH₃, —OCH₂CH₃,            —CH₂OCH₂CH₃, —OCH₂CH₂CH₃, and —CH₂OCH₂CH₂CH₃;-   ix) —[C(R^(37a))(R^(37b))]_(y)N(R^(25a))(R^(25b));    -   R^(25a) and R^(25b) are each independently chosen from:    -   a) —H;    -   b) —OR²⁶;        -   R²⁶ is hydrogen or C₁-C₄ linear alkyl;    -   c) substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   d) substituted or unsubstituted C₆ or C₁₀ aryl;    -   e) substituted or unsubstituted C₁-C₉ heterocyclic;    -   f) substituted or unsubstituted C₁-C₁₁ heteroaryl; or    -   g) R^(25a) and R^(25b) can be taken together to form a        substituted or unsubstituted ring having from 3 to 10 carbon        atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen,        and sulfur;        -   for example, —NH₂, —CH₂NH₂, —NHCH₃, —N(CH₃)₂, —NHOH,            —NHOCH₃, —NH(CH₂CH₃), —CH₂NHCH₃, —CH₂N(CH₃)₂,            —CH₂NH(CH₂CH₃), and the like;-   x) —[C(R^(37a))(R^(37b))]_(y)C(O)R²⁷;    -   R²⁷ is:    -   a) substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   b) —OR²⁸;        -   R²⁸ is hydrogen, substituted or unsubstituted C₁-C₄ linear            alkyl, substituted or unsubstituted C₆ or C₁₀ aryl,            substituted or unsubstituted C₁-C₉ heterocyclic, substituted            or unsubstituted C₁-C₁₁ heteroaryl;    -   c) —N(R^(29a))(R^(29b));        -   R^(29a) and R^(29b) are each independently hydrogen,            substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched,            or C₃-C₁₂ cyclic alkyl; substituted or unsubstituted C₆ or            C₁₀ aryl, substituted or unsubstituted C₁-C₉ heterocyclic,            substituted or unsubstituted C₁-C₁₁ heteroaryl; or R^(29a)            and R^(29b) can be taken together to form a substituted or            unsubstituted ring having from 3 to 10 carbon atoms and from            0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;        -   for example, —COCH₃, —CH₂COCH₃, —OCH₂CH₃, —CH₂COCH₂CH₃,            —COCH₂CH₂CH₃, —CH₂COCH₂CH₂CH₃, and the like;-   xi) —[C(R^(37a))(R^(37b))]_(y)OC(O)R³⁰;    -   R³⁰ is:    -   a) C₁-C₁₂ substituted or unsubstituted linear, branched, or        cyclic alkyl;    -   b) —N(R^(31a))(R^(31b));        -   R^(31a) and R^(31b) are each independently hydrogen,            substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched,            or C₃-C₁₂ cyclic alkyl; substituted or unsubstituted C₆ or            C₁₀ aryl, substituted or unsubstituted C₁-C₉ heterocyclic,            substituted or unsubstituted C₁-C₁₁ heteroaryl; or R^(31a)            and R^(31b) can be taken together to form a substituted or            unsubstituted ring having from 3 to 10 carbon atoms and from            0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;        -   for example, —OC(O)CH₃, —CH₂OC(O)CH₃, —OC(O)NH₂,            —CH₂OC(O)NH₂, —OC(O)NHCH₃, —CH₂OC(O)NHCH₃, —OC(O)N(CH₃)₂,            —CH₂OC(O)N(CH₃)₂, and the like;-   xii) —[C(R^(37a))(R^(37b))]_(y)NR³²C(O)R³³;    -   R³² is:    -   a) —H; or    -   b) substituted or unsubstituted C₁-C₄ linear, C₃-C₄ branched, or        C₃-C₄ cyclic alkyl;    -   R³³ is:    -   a) substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   b) —N(R^(34a))(R^(34b));        -   R^(34a) and R^(34b) are each independently hydrogen,            substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched,            or C₃-C₁₂ cyclic alkyl; substituted or unsubstituted C₆ or            C₁₀ aryl, substituted or unsubstituted C₁-C₉ heterocyclic,            substituted or unsubstituted C₁-C₁₁ heteroaryl; C₁-C₁₁            substituted or unsubstituted heteroaryl; or R^(34a) and            R^(34b) can be taken together to form a substituted or            unsubstituted ring having from 3 to 10 carbon atoms and from            0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;        -   for example, —NHC(O)CH₃, —CH₂NHC(O)CH₃, —NHC(O)NH₂,            —CH₂NHC(O)NH₂, —NHC(O)NHCH₃, —CH₂NHC(O)NHCH₃, —OC(O)N(CH₃)₂,            —CH₂NHC(O)N(CH₃)₂, and the like;-   xiii) —[C(R^(37a))(R^(37b))]_(y)CN; for example; —CN, —CH₂CN, and    —CH₂CH₂CN;-   xiv) —[C(R^(37a))(R^(37b))]_(y)NO₂; for example; —NO₂, —CH₂NO₂, and    —CH₂CH₂NO₂;-   xv) —[C(R^(37a))(R^(37b))]_(y)R³⁵; for example, —CH₂F, —CHF₂, —CF₃,    —CCl₃, or —CBr₃;    -   R³⁵ is C₁-C₁₀ linear, C₃-C₁₀ branched, or C₃-C₁₀ cyclic alkyl        substituted by from 1 to 21 halogen atoms chosen from —F, —Cl,        —Br, or —I;-   xvi) —[C(R^(37a))(R^(37b))]_(y)SO₂R³⁶;    -   R³⁶ is hydrogen, hydroxyl, substituted or unsubstituted C₁-C₄        linear or C₃-C₄ branched alkyl; substituted or unsubstituted C₆,        C₁₀, or C₁₋₄ aryl; C₇-C₁₅ alkylenearyl; substituted or        unsubstituted C₁-C₉ heterocyclic; or substituted or        unsubstituted C₁-C₁₁ heteroaryl;    -   for example, —SO₂H, —CH₂SO₂H, —SO₂CH₃, —CH₂SO₂CH₃, —SO₂C₆H₅, and        —CH₂SO₂C₆H₅; and-   xv) two hydrogen atoms on a ring carbon atom can be substituted to    form a ═O, ═S, or ═NH unit;    R^(37a) and R^(37b) are each independently hydrogen or C₁-C₄ alkyl;    and    the index y is an integer from 0 to 5.

A first embodiment of this aspect relates to compounds wherein R¹ and R²are taken together to form a 5-member substituted or unsubstituted C₁-C₄heterocyclic or a substituted or unsubstituted C₁-C₄ heteroaryl ring,non-limiting examples of which include a ring chosen from:

A first iteration of this embodiment relates to HIF-1α prolylhydroxylase inhibitors having the formula:

R²⁰⁰ represents from 0 to 2 substitutions for a ring hydrogen, whereinthe substitutions for hydrogen are independently chosen from:

i) C₁-C₄ linear or C₃-C₄ branched alkyl;

ii) C₁-C₄ linear or C₃-C₄ branched alkoxy;

iii) hydroxyl;

iv) cyano;

v) nitro;

vi) amino, methylamino, or dimethylamino;

vii) carboxy, methyl carboxy; or ethyl carboxy;

viii) formyl, acetyl, or propionyl;

ix) amido, methyl amido, or dimethyl amido;

x) halogen;

xi) heterocyclic; or

xii) heteroaryl.

Non-limiting examples of this iteration include HIF-1α prolylhydroxylase inhibitors having the formula:

A further iteration of this embodiment relates to HIF-1α prolylhydroxylase inhibitors wherein R¹ and R² are taken together to form a5-member substituted or unsubstituted heterocyclic or heteroaryl ringhaving more than one heteroatom in the ring. Non-limiting examplesinclude:

Another embodiment of this aspect relates to HIF-1α prolyl hydroxylaseinhibitors wherein R¹ and R² are taken together to form a substituted orunsubstituted C₄-C₁₁ heterocyclic or a substituted or unsubstitutedC₄-C₁₁ heteroaryl ring, non-limiting examples of which are chosen from:

Non-limiting examples of this embodiment include:

Another category of compounds has the formula:

wherein R²⁰⁰ and the index w are the same as defined herein above. Rrepresents from 0 to 5 substitutions for hydrogen, wherein each R isindependently chosen from:

-   i) C₁-C₁₂ substituted or unsubstituted linear, branched, or cyclic    alkyl; for example, methyl (C₁), (C₁), chloromethyl (C₁),    trifluoromethyl (C₁), aminomethyl (C₁), ethyl (C₂), hydroxymethyl    1-chloroethyl (C₂), 2-hydroxyethyl (C₂), 1,2-difluoroethyl (C₂),    n-propyl (C₃), iso-propyl (C₃), 3-carboxypropyl (C₃), cyclopropyl    (C₃), 2-methyl-cyclopropyl (C₃), n-butyl (C₄), sec-butyl (C₄),    iso-butyl (C₄), tert-butyl (C₄), cyclobutyl (C₄),    2,3-dihydroxycyclobutyl (C₄), pentyl (C₅), cyclopentyl (C₅), hexyl    (C₆), and cyclohexyl (C₆), and the like;-   ii) C₂-C₁₂ substituted or unsubstituted linear, branched, or cyclic    alkenyl; for example, ethenyl (C₂), 2-chloroethenyl (also    2-chlorovinyl) (C₂), 3-propenyl (C₃), 1-propenyl (also    2-methylethenyl) (C₃), isopropenyl (also 2-methylethen-2-yl) (C₃),    buten-4-yl (C₄), 4-hydroxybuten-1-yl (C₄), cyclobutenyl (C₄),    cyclopentenyl (C₅), cyclopentadienyl (C₅), cyclohexenyl (C₆),    7-hydroxy-7-methyloct-4-en-2-yl (C₉), and    7-hydroxy-7-methyloct-3,5-dien-2-yl (C₉), and the like;-   iii) C₂-C₁₂ substituted or unsubstituted linear or branched alkynyl;    for example, ethynyl (C₂), prop-2-ynyl (also propargyl) (C₃),    propyn-1-yl (C₃), 2-methyl-hex-4-yn-1-yl (C₇);    5-hydroxy-5-methylhex-3-ynyl (C₇), 6-hydroxy-6-methylhept-3-yn-2-yl    (C₈), 5-hydroxy-5-ethylhept-3-ynyl (C₉), and the like;-   iv) C₆ or C₁₀ substituted or unsubstituted aryl; for example, phenyl    (C₆), naphthylen-1-yl (C₁₀), naphthylen-2-yl (C₁₀), 4-fluorophenyl    (C₆), 2-hydroxyphenyl (C₆), 3-methylphenyl (C₆),    2-amino-4-fluorophenyl (C₆), 2-(N,N-diethylamino)phenyl (C₆),    2-cyanophenyl (C₆), 2,6-di-tert-butylphenyl (C₆), 3-methoxyphenyl    (C₆), 8-hydroxynaphthylen-2-yl (C₁₀), 4,5-dimethoxynaphthylen-1-yl    (C₁₀), 6-cyano-naphthylen-1-yl (C₁₀), and the like;-   v) C₁-C₉ substituted or unsubstituted heterocyclic; for example,    diazirinyl (C₁), aziridinyl (C₂), urazolyl (C₂), azetidinyl (C₃),    pyrazolidinyl (C₃), imidazolidinyl (C₃), oxazolidinyl (C₃),    isoxazolinyl (C₃), isoxazolyl (C₃), thiazolidinyl (C₃), isothiazolyl    (C₃), isothiazolinyl (C₃), oxathiazolidinonyl (C₃), oxazolidinonyl    (C₃), hydantoinyl (C₃), tetrahydropyranyl (C₄), pyrrolidinyl (C₄),    morpholinyl (C₄), piperazinyl (C₄), piperidinyl (C₄), dihydropyranyl    (C₅), tetrahydropyranyl (C₅), piperidin-2-onyl (valerolactam) (C₅),    and the like;-   vi) C₁-C₁₁ substituted or unsubstituted heteroaryl; for example,    1,2,3,4-tetrazolyl (C₁), [1,2,3]triazolyl (C₂), [1,2,4]triazolyl    (C₂), triazinyl (C₃), thiazolyl (C₃), 1H-imidazolyl (C₃), oxazolyl    (C₃), furanyl (C₄), thiopheneyl (C₄), pyrimidinyl (C₄), pyridinyl    (C₅), and the like;-   vii) halogen; for example, —F, —Cl, —Br, or —I;-   viii) —[C(R^(23a))(R^(23b))]_(x)OR¹⁰;    -   R¹⁰ is chosen from:    -   a) —H;    -   b) C₁-C₁₂ substituted or unsubstituted linear, branched, or        cyclic alkyl;    -   c) C₆ or C₁₀ substituted or unsubstituted aryl or alkylenearyl;    -   d) C₁-C₉ substituted or unsubstituted heterocyclic;    -   e) C₁-C₁₁ substituted or unsubstituted heteroaryl;        -   for example, —OH, —CH₂OH, —OCH₃, —CH₂OCH₃, —OCH₂CH₃,            —CH₂OCH₂CH₃, —OCH₂CH₂CH₃, and —CH₂OCH₂CH₂CH₃;-   ix) —[C(R^(23a))(R^(23b))]_(x)N(R^(11a))R^(11b));    -   R^(11a) and R^(11b) are each independently chosen from:    -   a) —H;    -   b) —OR¹²;        -   R¹² is hydrogen or C₁-C₄ linear alkyl;    -   c) C₁-C₁₂ substituted or unsubstituted linear, branched, or        cyclic alkyl;    -   d) C₆ or C₁₀ substituted or unsubstituted aryl;    -   e) C₁-C₉ substituted or unsubstituted heterocyclic;    -   f) C₁-C₁₁ substituted or unsubstituted heteroaryl; or    -   g) R^(11a) and R^(11b) can be taken together to form a        substituted or unsubstituted ring having from 3 to 10 carbon        atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen,        and sulfur;        -   for example, —NH₂, —CH₂NH₂, —NHCH₃, —N(CH₃)₂, —NHOH,            —NHOCH₃, —NH(CH₂CH₃), —CH₂NHCH₃, —CH₂N(CH₃)₂,            —CH₂NH(CH₂CH₃), and the like;-   x) —[C(R^(23a))(R^(23b))]_(x)C(O)R¹³    -   R¹³ is:    -   a) C₁-C₁₂ substituted or unsubstituted linear, branched, or        cyclic alkyl;    -   b) —OR¹⁴;        -   R¹⁴ is hydrogen, substituted or unsubstituted C₁-C₄ linear            alkyl, C₆ or C₁₀ substituted or unsubstituted aryl, C₁-C₉            substituted or unsubstituted heterocyclic, C₁-C₁₁            substituted or unsubstituted heteroaryl;    -   c) —N(R^(15a))(R^(15b));        -   R^(15a) and R^(15b) are each independently hydrogen, C₁-C₁₂            substituted or unsubstituted linear, branched, or cyclic            alkyl; C₆ or C₁₀ substituted or unsubstituted aryl; C₁-C₉            substituted or unsubstituted heterocyclic; C₁-C₁₁            substituted or unsubstituted heteroaryl; or R^(15a) and            R^(15b) can be taken together to form a substituted or            unsubstituted ring having from 3 to 10 carbon atoms and from            0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;            for example, —COCH₃, —CH₂COCH₃, —OCH₂CH₃, —CH₂COCH₂CH₃,            —COCH₂CH₂CH₃, —CH₂COCH₂CH₂CH₃, and the like;-   xi) —[C(R^(23a))(R^(23b))]_(x)OC(O)R¹⁶;    -   R¹⁶ is:    -   a) C₁-C₁₂ substituted or unsubstituted linear, branched, or        cyclic alkyl;    -   b) —N(R^(17a))(R^(17b));        -   R^(17a) and R^(1m) are each independently hydrogen, C₁-C₁₂            substituted or unsubstituted linear, branched, or cyclic            alkyl; C₆ or C₁₀ substituted or unsubstituted aryl; C₁-C₉            substituted or unsubstituted heterocyclic; C₁-C₁₁            substituted or unsubstituted heteroaryl; or R^(17a) and            R^(17b) can be taken together to form a substituted or            unsubstituted ring having from 3 to 10 carbon atoms and from            0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;-   xii) —[C(R^(23a))(R^(23b))]_(x)NR¹⁸C(O)R¹⁹;    -   R¹⁸ is:    -   a) —H; or    -   b) C₁-C₄ substituted or unsubstituted linear, branched, or        cyclic alkyl;

R¹⁹ is:

-   -   a) C₁-C₁₂ substituted or unsubstituted linear, branched, or        cyclic alkyl;    -   b) —N(R^(20a))(R^(20b));        -   R^(20a) and R^(20b) are each independently hydrogen, C₁-C₁₂            substituted or unsubstituted linear, branched, or cyclic            alkyl; C₆ or C₁₀ substituted or unsubstituted aryl; C₁-C₉            substituted or unsubstituted heterocyclic; C₁-C₁₁            substituted or unsubstituted heteroaryl; or R^(20a) and            R^(20b) can be taken together to form a substituted or            unsubstituted ring having from 3 to 10 carbon atoms and from            0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;            for example, —NHC(O)CH₃, —CH₂NHC(O)CH₃, —NHC(O)NH₂,            —CH₂NHC(O)NH₂, —NHC(O)NHCH₃, —CH₂NHC(O)NHCH₃, —OC(O)N(CH₃)₂,            —CH₂NHC(O)N(CH₃)₂, and the like;

-   xiii) —[C(R^(23a))(R^(23b))]_(x)CN; for example; —CN, —CH₂CN, and    —CH₂CH₂CN;

-   xiv) —[C(R^(23a))(R^(23b))]_(x)NO₂; for example; —NO₂, —CH₂NO₂, and    —CH₂CH₂NO₂;

-   xv) —[C(R^(23a))(R^(23b))]_(x)R²¹; for example, —CH₂F, —CHF₂, —CF₃,    —CCl₃, or —CBr₃;    -   R²¹ is C₁-C₁₀ linear, branched, or cyclic alkyl substituted by        from 1 to 21 halogen atoms chosen from —F, —Cl, —Br, or —I;

-   xvi) —[C(R^(23a))(R^(23b))]_(x)SO₂R²²;    -   R²² is hydrogen, hydroxyl, substituted or unsubstituted C₁-C₄        linear or branched alkyl; substituted or unsubstituted C₆, C₁₀,        or C₁₋₄ aryl; C₇-C₁₅ alkylenearyl; C₁-C₉ substituted or        unsubstituted heterocyclic; or C₁-C₁₁ substituted or        unsubstituted heteroaryl; for example, —SO₂H, —CH₂SO₂H, —SO₂CH₃,        —CH₂SO₂CH₃, —SO₂C₆H₅, and —CH₂SO₂C₆H₅;        R^(23a) and R^(23b) are each independently hydrogen or C₁-C₄        alkyl; and        the index x is an integer from 0 to 5.

Non-limiting examples of this category include compounds having theformula:

A further category of compounds relates to unsubstitutedN-benzyl-4-aminomethyl-3-hydroxypyridin-2-(1H)-ones having the formula:

wherein R¹ and R² are each independently chosen from:

i) hydrogen;

ii) substituted or unsubstituted C₁-C₁₀ linear, branched, or cyclicalkyl;

iii) substituted or unsubstituted C₂-C₁₀ linear, branched, or cyclicalkenyl;

iv) substituted or unsubstituted C₂-C₁₀ linear or branched alkynyl;

v) substituted or unsubstituted C₆ or C₁₀ aryl;

vi) substituted or unsubstituted C₁-C₉ heterocyclic; or

vii) substituted or unsubstituted C₁-C₉ heteroaryl.

The first aspect of this category relates to HIF-1α prolyl hydroxylaseinhibitors wherein R² is hydrogen and R¹ is substituted or unsubstitutedC₁-C₉ heterocyclic or C₁-C₉ heteroaryl. In a first embodiment, R¹ is asubstituted heterocyclic group, non-limiting examples of which includeaziridinyl (C₂), azetidinyl (C₃), pyrrolidinyl (C₄), morpholinyl (C₄),piperazinyl (C₄), piperidinyl (C₄), piperidin-2-onyl (valerolactam)(C₅), and azepan-2-only (caprolactam) (C₆), wherein the R¹ unit can bebonded to the nitrogen atom at any position in the ring. In addition,the C₁-C₉ heterocyclic or C₁-C₉ heteroaryl ring can be substituted atany position whether a ring carbon or a ring heteroatom, for example, aring nitrogen. Non-limiting examples of this embodiment include:

In another embodiment, R² is hydrogen and R¹ is substituted orunsubstituted C₃-C₁₂ cycloalkyl wherein the cycloalkyl ring can besubstituted at any ring position. Non-limiting examples of thisembodiment include:

A yet further category of compounds relates to unsubstitutedN-benzyl-4-aminomethyl-3-hydroxypyridin-2-(1H)-ones having the formula:

R¹ and R² are each independently hydrogen or substituted orunsubstituted C₁-C₁₀ linear or branched alkyl, wherein the alkyl unitcan be substituted by one or more units independently chosen from:

i) C₁-C₈ linear, branched, or cyclic alkoxy;

ii) hydroxy;

iii) halogen;

iv) cyano;

v) amino, C₁-C₈ mono-alkylamino, C₁-C₈ di-alkylamino;

vi) —SR⁴⁰; R⁴⁰ is hydrogen or C₁-C₄ linear or branched alkyl;

vii) substituted or unsubstituted C₆ of C₁₀ aryl;

viii) substituted or unsubstituted C₁-C₉ heterocyclic; or

ix) substituted or unsubstituted C₁-C₉ heteroaryl.

Non-limiting examples of this category include:

A still further category of the disclosed compounds has the formula:

wherein R²⁰⁰ and the index w are the same as defined herein above. Rrepresents from 0 to 5 substitutions for hydrogen, wherein each R isindependently chosen from:

-   i) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched, or    C₃-C₁₂ cyclic alkyl; for example, methyl (C₁), (C₁), chloromethyl    (C₁), trifluoromethyl (C₁), aminomethyl (C₁), ethyl (C₂),    hydroxymethyl 1-chloroethyl (C₂), 2-hydroxyethyl (C₂),    1,2-difluoroethyl (C₂), n-propyl (C₃), iso-propyl (C₃),    3-carboxypropyl (C₃), cyclopropyl (C₃), 2-methyl-cyclopropyl (C₃),    n-butyl (C₄), sec-butyl (C₄), iso-butyl (C₄), tert-butyl (C₄),    cyclobutyl (C₄), 2,3-dihydroxycyclobutyl (C₄), pentyl (C₅),    cyclopentyl (C₅), hexyl (C₆), and cyclohexyl (C₆), and the like;-   ii) substituted or unsubstituted C₂-C₁₂ linear, C₃-C₁₂ branched, or    C₃-C₁₂ cyclic alkenyl; for example, ethenyl (C₂), 2-chloroethenyl    (also 2-chlorovinyl) (C₂), 3-propenyl (C₃), 1-propenyl (also    2-methylethenyl) (C₃), isopropenyl (also 2-methylethen-2-yl) (C₃),    buten-4-yl (C₄), 4-hydroxybuten-1-yl (C₄), cyclobutenyl (C₄),    cyclopentenyl (C₅), cyclopentadienyl (C₅), cyclohexenyl (C₆),    7-hydroxy-7-methyloct-4-en-2-yl (C₉), and    7-hydroxy-7-methyloct-3,5-dien-2-yl (C₉), and the like;-   iii) substituted or unsubstituted C₂-C₁₂ linear or C₃-C₁₂ branched    alkynyl; for example, ethynyl (C₂), prop-2-ynyl (also propargyl)    (C₃), propyn-1-yl (C₃), 2-methyl-hex-4-yn-1-yl (C₇);    5-hydroxy-5-methylhex-3-ynyl (C₇), 6-hydroxy-6-methylhept-3-yn-2-yl    (C₈), 5-hydroxy-5-ethylhept-3-ynyl (C₉), and the like;-   iv) substituted or unsubstituted C₆ or C₁₀ aryl; for example, phenyl    (C₆), naphthylen-1-yl (C₁₀), naphthylen-2-yl (C₁₀), 4-fluorophenyl    (C₆), 2-hydroxyphenyl (C₆), 3-methylphenyl (C₆),    2-amino-4-fluorophenyl (C₆), 2-(N,N-diethylamino)phenyl (C₆),    2-cyanophenyl (C₆), 2,6-di-tert-butylphenyl (C₆), 3-methoxyphenyl    (C₆), 8-hydroxynaphthylen-2-yl (C₁₀), 4,5-dimethoxynaphthylen-1-yl    (C₁₀), 6-cyano-naphthylen-1-yl (C₁₀), and the like;-   v) substituted or unsubstituted C₁-C₉ heterocyclic; for example,    diazirinyl (C₁), aziridinyl (C₂), urazolyl (C₂), azetidinyl (C₃),    pyrazolidinyl (C₃), imidazolidinyl (C₃), oxazolidinyl (C₃),    isoxazolinyl (C₃), isoxazolyl (C₃), thiazolidinyl (C₃), isothiazolyl    (C₃), isothiazolinyl (C₃), oxathiazolidinonyl (C₃), oxazolidinonyl    (C₃), hydantoinyl (C₃), tetrahydropyranyl (C₄), pyrrolidinyl (C₄),    morpholinyl (C₄), piperazinyl (C₄), piperidinyl (C₄), dihydropyranyl    (C₅), tetrahydropyranyl (C₅), piperidin-2-onyl (valerolactam) (C₅),    and the like;-   vi) substituted or unsubstituted C₁-C₁₁ heteroaryl; for example,    1,2,3,4-tetrazolyl (C₁), [1,2,3]triazolyl (C₂), [1,2,4]triazolyl    (C₂), triazinyl (C₃), thiazolyl (C₃), 1H-imidazolyl (C₃), oxazolyl    (C₃), furanyl (C₄), thiopheneyl (C₄), pyrimidinyl (C₄), pyridinyl    (C₅), and the like;-   vii) halogen; for example, —F, —Cl, —Br, or —I;-   viii) —[C(R^(23a))(R^(23b))]_(x)OR¹⁰;    -   R¹⁰ is chosen from:    -   a) —H;    -   b) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   c) substituted or unsubstituted C₆ or C₁₀ aryl or C₇ or C₁₀        alkylenearyl;    -   d) substituted or unsubstituted C₁-C₉ heterocyclic;    -   e) substituted or unsubstituted C₁-C₁₁ heteroaryl;        -   for example, —OH, —CH₂OH, —OCH₃, —CH₂OCH₃, —OCH₂CH₃,            —CH₂OCH₂CH₃, —OCH₂CH₂CH₃, and —CH₂OCH₂CH₂CH₃;-   ix) —[C(R^(23a))(R^(23b))]_(x)N(R^(11a))(R^(11b));    -   R^(11a) and R^(11b) are each independently chosen from:    -   a) —H;    -   b) —OR¹²;        -   R¹² is hydrogen or C₁-C₄ linear alkyl;    -   c) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   d) substituted or unsubstituted C₆ or C₁₀ aryl;    -   e) substituted or unsubstituted C₁-C₉ heterocyclic;    -   f) substituted or unsubstituted C₁-C₁₁ heteroaryl; or    -   g) R^(11a) and R^(11b) can be taken together to form a        substituted or unsubstituted ring having from 3 to 10 carbon        atoms and from 0 to 3 heteroatoms chosen from oxygen, nitrogen,        and sulfur;        -   for example, —NH₂, —CH₂NH₂, —NHCH₃, —N(CH₃)₂, —NHOH,            —NHOCH₃, —NH(CH₂CH₃), —CH₂NHCH₃, —CH₂N(CH₃)₂,            —CH₂NH(CH₂CH₃), and the like;-   x) —[C(R^(23a))(R^(23b))]_(x)C(O)R¹³;    -   R¹³ is:    -   a) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   b) —OR¹⁴;        -   R¹⁴ is hydrogen, substituted or unsubstituted C₁-C₄ linear            alkyl, substituted or unsubstituted C₆ or C₁₀ aryl,            substituted or unsubstituted C₁-C₉ heterocyclic, substituted            or unsubstituted C₁-C₁₁ heteroaryl;    -   c) —N(R^(15a))(R^(15b));        -   R^(15a) and R^(15b) are each independently hydrogen,            substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,            or C₃-C₁₂ cyclic alkyl; substituted or unsubstituted C₆ or            C₁₀ aryl; substituted or unsubstituted C₁-C₉ heterocyclic;            substituted or unsubstituted C₁-C₁₁ heteroaryl; or R^(15a)            and R^(15b) can be taken together to form a substituted or            unsubstituted ring having from 3 to 10 carbon atoms and from            0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;        -   for example, —COCH₃, —CH₂COCH₃, —OCH₂CH₃, —CH₂COCH₂CH₃,            —COCH₂CH₂CH₃, —CH₂COCH₂CH₂CH₃, and the like;-   xi) —[C(R^(23a))(R^(23b))]_(x)OC(O)R¹⁶;    -   R¹⁶ is:    -   a) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   b) —N(R^(17a))(R^(17b));        -   R^(17a) and R^(17b) are each independently hydrogen,            substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,            or C₃-C₁₂ cyclic alkyl; substituted or unsubstituted C₆ or            C₁₀ aryl; substituted or unsubstituted C₁-C₉ heterocyclic;            substituted or unsubstituted C₁-C₁₁ heteroaryl; or R^(17a)            and R^(1m) can be taken together to form a substituted or            unsubstituted ring having from 3 to 10 carbon atoms and from            0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;-   xii) —[C(R^(23a))(R^(23b))]_(x)NR¹⁸C(O)R¹⁹;    -   R¹⁸ is:    -   a) —H; or    -   b) substituted or unsubstituted C₁-C₄ linear, C₃-C₄ branched, or        C₃-C₄ cyclic alkyl;

R¹⁹ is:

-   -   a) substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,        or C₃-C₁₂ cyclic alkyl;    -   b) —N(R^(20a))(R^(20b));        -   R^(20a) and R^(20b) are each independently hydrogen,            substituted or unsubstituted C₁-C₁₂ linear, C₃-C₁₂ branched,            or C₃-C₁₂ cyclic alkyl; substituted or unsubstituted C₆ or            C₁₀ aryl; substituted or unsubstituted C₁-C₉ heterocyclic;            substituted or unsubstituted C₁-C₁₁ heteroaryl; or R^(20a)            and R^(20b) can be taken together to form a substituted or            unsubstituted ring having from 3 to 10 carbon atoms and from            0 to 3 heteroatoms chosen from oxygen, nitrogen, and sulfur;        -   for example, —NHC(O)CH₃, —CH₂NHC(O)CH₃, —NHC(O)NH₂,            —CH₂NHC(O)NH₂, —NHC(O)NHCH₃, —CH₂NHC(O)NHCH₃, —OC(O)N(CH₃)₂,            —CH₂NHC(O)N(CH₃)₂, and the like;

-   xiii) —[C(R^(23a))(R^(23b))]_(x)CN; for example; —CN, —CH₂CN, and    —CH₂CH₂CN;

-   xiv) —[C(R^(23a))(R²³)]_(x)NO₂; for example; —NO₂, —CH₂NO₂, and    —CH₂CH₂NO₂;

-   xv) —[C(R^(23a))(R^(23b))]_(x)R²¹; for example, —CH₂F, —CHF₂, —CF₃,    —CCl₃, or —CBr₃;    -   R²¹ is C₁-C₁₀ linear, branched, or cyclic alkyl substituted by        from 1 to 21 halogen atoms chosen from —F, —Cl, —Br, or —I;

-   xvi) —[C(R^(23a))(R^(23b))]_(x)SO₂R²²;    -   R²² is hydrogen, hydroxyl, substituted or unsubstituted C₁-C₄        linear or C₃-C₄ branched alkyl; substituted or unsubstituted C₆,        C₁₀, or C₁₋₄ aryl; C₇-C₁₅ alkylenearyl; substituted or        unsubstituted C₁-C₉ heterocyclic; or substituted or        unsubstituted C₁-C₁₁ heteroaryl; for example, —SO₂H, —CH₂SO₂H,        —SO₂CH₃, —CH₂SO₂CH₃, —SO₂C₆H₅, and —CH₂SO₂C₆H₅;        R^(23a) and R^(23b) are each independently hydrogen or C₁-C₄        alkyl; and        the index x is an integer from 0 to 5.

One aspect embodiment of this category relates to HIF-1α prolylhydroxylase inhibitors wherein R¹ and R² are taken together to form a5-member substituted or unsubstituted C₁-C₄ heterocyclic or asubstituted or unsubstituted C₁-C₄ heteroaryl ring, non-limitingexamples of which include a ring chosen from:

Another aspect of this category relates to HIF-1α prolyl hydroxylaseinhibitors wherein R¹ and R² are taken together to form a substituted orunsubstituted C₄-C₁₁ heterocyclic or a substituted or unsubstitutedC₄-C₁₁ heteroaryl ring, non-limiting examples of which are chosen from:

Non-limiting examples of this category include compounds having theformula:

A further category of the disclosed compounds has the formula:

wherein R represents from 1 to 5 optional substitutions for a phenylring hydrogen atom, R¹ and R² are each independently hydrogen orsubstituted or unsubstituted C₁-C₁₀ linear or branched alkyl, whereinthe alkyl unit can be substituted by one or more units independentlychosen from:

i) C₁-C₈ linear, C₃-C₈ branched, or C₃-C₈ cyclic alkoxy;

ii) hydroxy;

iii) halogen;

iv) cyano;

v) amino, C₁-C₈ mono-alkylamino, C₁-C₈ di-alkylamino;

vi) —SR⁴⁰; R⁴⁰ is hydrogen or C₁-C₄ linear or branched alkyl;

vii) substituted or unsubstituted C₆ of C₁₀ aryl;

viii) substituted or unsubstituted C₁-C₉ heterocyclic; or

ix) substituted or unsubstituted C₁-C₉ heteroaryl.

Non-limiting examples of this category include:

A still yet further category of the disclosed HIF-1α prolyl hydroxylaseinhibitors relates to compounds having the formula:

wherein R¹ and R² are taken together to form a substituted orunsubstituted piperazine ring, the substitutions on the ring as definedfor R²⁰⁰ herein above.

A yet still further category of the disclosed HIF-1α prolyl hydroxylaseinhibitors have the formula:

wherein R¹ and R² can be taken together to form a substituted orunsubstituted heterocyclic or heteroaryl ring having from 2 to 20 carbonatoms and from 1 to 7 heteroatoms wherein the rings formed exclude apiperazine ring.

Also disclosed herein are N-substituted benzyl or N-substitutedsulfonylaryl-3-hydroxypyridin-2-(1H)-ones having the formula:

that can be used to stimulate the cellular immune response in a subject.For these compounds, Z and L are the same as disclosed herein above.Non-limiting examples of these compounds include:

-   1-(4-chlorobenzyl)-3-hydroxypyridin-2(1H)-one having the formula:

-   1-(3-chlorobenzyl)-3-hydroxypyridin-2(1H)-one having the formula:

and

-   1-(2-chlorobenzyl)-3-hydroxypyridin-2(1H)-one having the formula:

Further disclosed herein are N-substituted benzyl or N-substitutedsulfonylaryl-5-substituted-3-hydroxypyridin-2-(1H)-ones having theformula:

wherein Y is substituted or unsubstituted phenyl, Z and L are the sameas defined herein above.

One aspect of Y relates to a phenyl group that is substituted with from1 to 5 halogen atoms, for example, Y is chosen from 2-chlorophenyl,3-chlorophenyl, 2-fluorophenyl, 3-fluorophenyl, or 4-fluorophenyl. Afurther aspect of Y units relates to compounds wherein Y is chosen from2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl,2,6-difluorophenyl, 2,3-dichlorophenyl, 2,4-dichlorophenyl,2,5-dichlorophenyl, and 2,6-dichlorophenyl.

A non-limiting example of compounds according to this category include1-(4-chlorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one havingthe formula:

Further non-limiting examples include:

-   1-(2-chlorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(2-chlorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(2-chlorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-chlorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-chlorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-chlorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(4-chlorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(4-chlorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(2-fluorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(2-fluorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(2-fluorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-fluorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-fluorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-fluorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(4-fluorobenzyl)-5-(2-chlorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(4-fluorobenzyl)-5-(3-chlorophenyl)-3-hydroxypyridin-2(1H)-one-   1-(4-fluorobenzyl)-5-(4-chlorophenyl)-3-hydroxypyridin-2(1H)-one-   1-(2-chlorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(2-chlorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(2-chlorobenzyl)-5-(4-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-chlorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-chlorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-chlorobenzyl)-5-(4-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(4-chlorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(4-chlorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(4-chlorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one-   1-(2-fluorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(2-fluorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(2-fluorobenzyl)-5-(4-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-fluorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-fluorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(3-fluorobenzyl)-5-(4-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(4-fluorobenzyl)-5-(2-fluorophenyl)-3-hydroxypyridin-2(1H)-one;-   1-(4-fluorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one;    and-   1-(4-fluorobenzyl)-5-(3-fluorophenyl)-3-hydroxypyridin-2(1H)-one.

The disclosed compounds are organized into several categories for thestrictly non-limiting purpose of describing alternatives for syntheticstrategies for the preparation of subgenera of compounds within thescope of the disclosed compounds that are not expressly exemplifiedherein. This mental organization into categories does not imply anythingwith respect to increased or decreased biological efficacy with respectto any of the compounds or compositions of matter described herein.

Category I of the disclosed HIF-1α prolyl hydroxylase inhibitors relatesto compounds having the formula:

wherein A is a substituted or unsubstituted heterocyclic or heteroarylring having from 2 to 20 carbon atoms and from 1 to 7 heteroatoms, R²⁰⁰represents from 0 to 40 substitutions form hydrogen, R represents from 1to 5 substitutions for hydrogen as defined herein above, and the index nis from 1 to 5. Table I provides representative examples of compoundsaccording to this category.

TABLE I No. R A ring A1 3-methoxy pyrrolidin-1-yl A2 3-methoxy3-hydroxypyrrolidin-1-yl A3 3-methoxy 2-(pyrdin-2-yl)pyrrolidin-1-yl A43-methoxy 2-methylcarboxypyrrolidin-1-yl A5 3-methoxy2-(methoxymethyl)pyrrolidin-1-yl A6 3-methoxy thiazolidin-3-yl A73-methoxy 1H-imidazol-1-yl A8 3-methoxy piperidin-1-yl A9 3-methoxy4-benzylpiperidin-1-yl A10 3-methoxy 1,4′-bipiperidinyl-1′-yl A113-methoxy piperazin-1-yl A12 3-methoxy 4-benzylpiperazin-1-yl A133-methoxy 4-(2-methoxyphenyl)piperazin-1-ylmethyl A14 3-methoxy4-(6-chloropyridazin-3-yl)piperazin-1-yl A15 3-methoxy1,4-dioxa-8-azaspiro[4,5]dec-8-yl A16 3-methoxy morpholin-4-yl A173-methoxy thiomorpholin-4-yl A18 3-methoxy azepan-1-yl A19 3-methoxyazocan-1-yl A20 3-methoxy 3,4-dihydroquinolin-1(2H)-yl A21 4-chloropyrrolidin-1-yl A22 4-chloro 3-hydroxypyrrolidin-1-yl A23 4-chloro2-(pyrdin-2-yl)pyrrolidin-1-yl A24 4-chloro2-methylcarboxypyrrolidin-1-yl A25 4-chloro2-(methoxymethyl)pyrrolidin-1-yl A26 4-chloro thiazolidin-3-yl A274-chloro 1H-imidazol-1-yl A28 4-chloro piperidin-1-yl A29 4-chloro4-benzylpiperidin-1-yl A30 4-chloro 1,4′-bipiperidinyl-1′-yl A314-chloro piperazin-1-yl A32 4-chloro 4-benzylpiperazin-1-yl A33 4-chloro4-(2-methoxyphenyl)piperazin-1-ylmethyl A34 4-chloro4-(6-chloropyridazin-3-yl)piperazin-1-yl A35 4-chloro1,4-dioxB-8-azaspiro[4,5]dec-8-yl A36 4-chloro morpholin-4-yl A374-chloro thiomorpholin-4-yl A38 4-chloro azepan-1-yl A39 4-chloroazocan-1-yl A40 4-chloro 3,4-dihydroquinolin-1(2H)-yl A41 4-chloro4-tert-butoxycarbonylpiperazin-1-yl

The disclosed compounds of this category can be prepared by theprocedure outlined herein below in Scheme I and described in Example 1.

Example 1tert-Butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate(4)

Preparation of 3-(tert-butyldimethylsilanyloxy)-1H-pyridin-2-one (1):3-Hydroxypyridin-2(1H)-one (15 g, 135 mmol) and imidazole (23 g, 338mmol) were suspended in dimethylformamide (200 mL) under inertatmosphere. A solution of ten-butyldimethylsilyl chloride (20.5 g, 136mmol) in dimethylformamide (200 mL) is added dropwise at roomtemperature over 30 minutes. The reaction was then allowed to stirovernight. The resulting solution was then poured into water (300 mL)and the mixture extracted with tert-butyl methyl ether (3×500 mL). Thecombined organic layer was washed with water (300 mL), brine (300 mL)then dried over Na₂SO₄. The solvent is removed under reduced pressureand the crude product crystallized from heptanes to afford 16.3 g (53%yield) of the desired product. ¹H NMR (250 MHz, CDCl₃) δ ppm 12.98 (1H,m); 6.91 (1H, dd, J=1. Hz, J=6.8 Hz); 6.81 (1H, dd, J=1.8 Hz, J=7.2 Hz);6.02-6.007 (1H, m); 0.90 (9H, s), and 0.17 (6H, s).

Preparation of3-(tert-butyldimethylsilanyloxy)-1-(3-chlorobenzyl)-1H-prydin-2-one (2):At 0° C. under an inert atmosphere, a solution of 4-chlorobenzylchloride (4.44 mmol) in THF (10 mL) was added dropwise to a solution of3-(tert-butyldimethylsilanyloxy)-1H-pyridin-2-one, 1, (1 g, 4.44 mmol)and CsCO₃ (2.17 g, 6.66 mmol) in THF (10 mL). The reaction solution wasallowed to warm to room temperature and stirring was continuedovernight. The resulting solution was diluted with water (40 mL) andthen extracted with EtOAc (3×30 mL). The combined organic layer waswashed with brine (30 mL) then dried over Na₂SO₄. The solvent is removedunder reduced pressure and the crude product purified over silica(EtOAc:heptane 4:1) to afford the desired product as a white solid.

Preparation of 1-(4-chlorobenzyl)-3-hydroxypyridin-2(1H)-one (3): To asolution of3-(tert-butyldimethylsilanyloxy)-1-(3-chlorobenzyl)-1H-prydin-2-one, 2,(2.36 g, 10 mmol) in EtOAc (25 mL) as added 5 M HCl (25 mL) withvigorous stirring at room temperature. The reaction was monitored by TLCfor the disappearance of starting material and was complete within 30minutes. The organic layer was decanted and the aqueous phase extractedwith dichloromethane (2×50 mL). The combined organic layers were driedover Na₂SO₄ and the solvent removed under reduced pressure. The crudeproduct was recrystallized from dichloromethane. The yield was nearlyquantitative. ¹H NMR (360 MHz, DMSO-d₆) δ ppm 5.12 (2H, s); 6.13 (1H, t,J=7.04); 6.71 (1H, dd, J=7.04, 1.59); 7.23-7.28 (2H, m); 7.36-7.43 (2H,m); 9.10 (1H, br. s).

Preparation oftert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydro-pyridin-4-yl]methyl}piperazine-1-carboxylate(4): tert-Butyl piperazine-1-carboxylate (97.6 mmol), formaldehyde (8 mLof a 37% soln., 97.6 mmol) and acetic acid (8 mL) were dissolved inethanol (350 mL) and the solution stirred for 1 hour at roomtemperature. A solution of1-(4-chlorobenzyl)-3-hydroxypyridin-2(1H)-one, 3, (48.8 mmol) in ethanol(350 mL) was added dropwise over 30 minutes. After 3 days of stirring,formaldehyde (3 mL) was added and the reaction heated to 50° C. afterwhich the reaction solution was concentrated under reduced pressure toapproximately 500 mL. The desired product is obtained by crystallizationfrom ethanol. ¹H NMR (250 MHz, CDCl₃) d ppm 1.46 (s, 9H); 2.38-2.57 (m,4H); 3.40-3.49 (m, 4H); 3.51 (s, 2H); 5.13 (s, 2H); 6.13 (d, J=7.16 Hz),1H); 6.79 (d, J=7.16 Hz, 1H); 7.20-7.41 (m, 4H); 8.33-8.85 (m, 1H).

Category II of the disclosed prolyl hydroxylase inhibitors relates tocompounds having the formula:

wherein A is a substituted or unsubstituted heterocyclic or heteroarylring having from 2 to 20 carbon atoms and from 1 to 7 heteroatoms, andR²⁰⁰ represents from 0 to 40 substitutions form hydrogen. Table IIprovides representative examples of compounds according to thiscategory.

TABLE II No. A ring B1 pyrrolidin-1-yl B2 3-hydroxypyrrolidin-1-yl B32-(pyrdin-2-yl)pyrrolidin-1-yl B4 2-methylcarboxypyrrolidin-1-yl B52-(methoxymethyl)pyrrolidin-1-yl B6 thiazolidin-3-yl B7 1H-imidazol-1-ylB8 piperidin-1-yl B9 4-benzylpiperidin-1-yl B10 1,4′-bipiperidinyl-1′-ylB11 piperazin-1-yl B12 4-benzylpiperazin-1-yl B134-(2-methoxyphenyl)piperazin-1-ylmethyl B144-(6-chloropyridazin-3-yl)piperazin-1-yl B151,4-dioxa-8-azaspiro[4,5]dec-8-yl B16 morpholin-4-yl B17thiomorpholin-4-yl B18 azepan-1-yl B19 azocan-1-yl B203,4-dihydroquinolin-1(2H)-yl

The compounds according to Category II can be prepared according to theprocedure outlined in Scheme I and disclosed in Example 1. The followingare further examples of inhibitors according to Category II.

1-Benzyl-3-hydroxy-4-(piperidin-1-ylmethyl)pyridin-2(1H)-one

¹H NMR (300 MHz, CD₃OD) δ 1.81 (m, 6H), 3.07 (m, 2H), 3.51 (m, 2H), 4.23(s, 2H), 5.24 (s, 2H), 6.31 (d, J=6.9 Hz, 1H), 7.35 (m, 6H); ¹⁹F NMR(252 MHz, CD₃OD) δ 85.5; 13C NMR (75 MHz, DMSO) δ 21.3, 22.7, 51.8,52.5, 53.1, 106.4, 117.4, 127.7, 128.0, 128.2, 128.9, 137.3, 147.4,158.0; ES MS (M+1) 299.12; HRMS Calcd. For C₁₈H₂₂N₂O₂, 298.38. Found(M+1) 299.17.

1-Benzyl-3-hydroxy-4-(morpholin-4-ylmethyl)pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 3.25 (m, 4H), 3.81 (m, 4H), 4.18 (s, 2H), 5.17(s, 2H), 6.31 (d, J=6.9 Hz, 1H), 7.35 (m, 6H); ¹⁹F NMR (300 MHz, DMSO) δ88.5; ¹³C NMR (300 MHz, DMSO) δ 51.6, 51.8, 53.4, 63.5, 107.9, 119.1,127.8, 128.0, 128.2, 128.9, 137.3, 147.5, 158.3; ES MS (M+1) 301.12;HRMS Calcd. For C₁₇H₂₀N₂O₃, 300.35.

1-Benzyl-3-hydroxy-4-(thiomorpholin-4-ylmethyl)pyridin-2(1H)-one

¹HNMR (300 MHz, DMSO) δ 2.92 (m, 4H), 3.38 (m, 4H), 4.17 (s, 2H), 5.16(s, 2H), 6.29 (d, J=7.5 Hz, 1H), 7.34 (m, 6H), 9.97 (s, 1H); ¹⁹F NMR(300 MHz, DMSO) δ 88.4; ¹³C NMR (75 MHz, DMSO) δ 24.3, 51.9, 53.4, 53.7,107.9, 110.9, 127.8, 128.0, 128.2, 128.8, 137.2, 147.6, 157.6; ES MS(M+1) 317.14; HRMS Calcd. For C₁₇H₂₀N₂O₂S, 316.42. Found: (M+1) 317.13.

1-Benzyl-3-hydroxy-4-(thiazolidin-3-ylmethyl)pyridin-2(1H)-one

¹HNMR (300 MHz, DMSO) δ 3.09 (t, J=6.3 Hz, 2H), 3.42 (t, J=6.3 Hz, 2H),4.03 (s, 2H), 4.29 (s, 2H), 5.16 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34(m, 6H), 10.48 (broad s, 1H); ¹⁹FNMR (300 MHz, DMSO) δ 87.9; ¹³CNMR (75MHz, DMSO) δ 28.3, 48.3, 50.1, 56.3, 57.0, 107.4, 122.1, 127.8, 128.2,128.8, 137.4, 146.3, 157.6; ES MS (M+1) 303.08; Anal. Calcd forC₁₈H₁₉N₂O₄SF, C, 51.92; H, 4.60; N, 6.73; S, 7.70. Found: C, 51.67; H,4.48; N, 6.69; S, 7.65.

1-Benzyl-3-hydroxy-4-(pyrrolidin-1-ylmethyl)pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.96 (s, 4H), 3.16 (s, 2H), 3.43 (s, 2H), 4.23(s, 4H), 5.17 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34 (m, 6H); ¹⁹F NMR(252 MHz, DMSO) δ 88.7; ¹³C NMR (75 MHz, DMSO) δ 22.8, 50.9, 51.8, 53.7,107.3, 118.0, 128.0, 128.2, 128.9, 137.3, 146.7, 157.6; ES MS (M+1)285.13; Anal. Calcd. For C₁₉H₂₁F₃N₂O₄, C, 57.28; H, 5.31; N, 7.03.Found: C, 57.10; H, 5.11; N, 7.02.

1-Benzyl-3-hydroxy-4-(4-benzylpiperidin-1-ylmethyl)pyridin-2(1H)-one

¹H NMR (DMSO) δ 1.43 (m, 2H), 1.72 (m, 4H), 2.96 (m, 2H), 3.41 (m, 3H),4.09 (s, 2H), 5.16 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.35 (m, 11H); ¹⁹FNMR (252 MHz, DMSO) δ 88.8; ¹³C NMR (75 MHz, DMSO) δ; ES MS (M+1)389.21; HRMS Calcd. For C₂₅H₂₈N₂O₂, 388.50. Found (M+1) 389.22.

1-Benzyl-3-hydroxy-4-(4-benzylpiperazin-1-ylmethyl)pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 3.11 (broad s, 4H), 3.81 (s, 2H), 4.18 (s, 2H),5.15 (s, 2H), 6.24 (d, J=7.2 Hz, 1H), 7.34 (m, 6H), 7.46 (m, 5H); ¹⁹FNMR (252 MHz, DMSO) δ 88.2; 13C (75 MHz, DMSO) δ; ES MS (M+1) 390.21;HRMS Calcd. For C₂₄H₂₇N₃O₂, 389.49. Found (M+1) 390.21.

1-Benzyl-3-hydroxy-4-[(3-hydroxypyrrolidin-1-yl)methyl]pyridin-2(1H)-one

¹HNMR (300 MHz, DMSO) δ 1.90 (m, 1H), 3.18 (m, 2H), 3.47 (m, 3H), 4.24(s, 2H), 4.43 (s, 1H), 5.17 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34 (m,6H); ¹⁹F NMR (252 MHz, DMSO) δ 89.0; ¹³C NMR (75 MHz, DMSO) δ 51.8,52.6, 61.3, 68.6, 107.4, 117.9, 128.0, 128.2, 128.9, 137.3, 146.7,157.6; ES MS (M+1) 301.13; HRMS Calcd. For C₁₇H₂₀N₂O₃, 300.35. Found:(M+1) 301.15.

1-Benzyl-3-hydroxy-4-(1,4-dioxa-8-azaspiro[4,5]dec-8-ylmethyl)pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.90 (m, 4H), 3.11 (m, 2H), 3.43 (m, 2H), 3.93(s, 4H), 4.19 (s, 2H), 5.16 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34 (m,6H), 10.01 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.3; ¹³C NMR (75MHz, DMSO) δ 31.7, 50.7, 51.9, 52.5, 64.5, 101.1, 108.0, 116.5, 127.8,128.0, 128.3, 128.9, 137.3, 147.5 157.6; ES MS (M+1) 357.19; HRMS Calcd.For C₂₀H₂₄N₄O₂, 356.42. Found (M+1) 357.18.

1-Benzyl-3-hydroxy-4-azepan-1-ylmethylpyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.61 (m, 4H), 1.80 (m, 4H), 3.20 (m, 4H), 4.17(s, 2H), 5.16 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34 (m, 6H); ¹⁹F NMR(252 MHz, DMSO) δ 88.9; ¹³C NMR (75 MHz, DMSO) δ 22.8, 26.4, 51.8, 53.4,54.4, 107.6, 117.2, 127.9, 128.0, 18.2, 128.9, 137.3, 147.2, 157.6; ESMS (M+1) 313.18; HRMS Calcd. For C₁₉H₂₄N₂O₄, 312.41. Found (M+1) 313.19.

1-Benzyl-3-hydroxy-4-(azocan-1-ylmethyl)pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.59 (m, 10H), 3.18 (m, 2H), 3.38 (m, 2H), 4.17(s, 2H), 5.16 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34 (m, 6H); ¹⁹F NMR(252 MHz, DMSO) δ 88.9; ¹³C NMR (75 MHz, DMSO) δ; ES MS (M+1) 327.2;HRMS Calcd. For C₂₀H₂₆N₂O₂, 326.43. Found (M+1) 327.20.

1-Benzyl-3-hydroxy-(1,4′-bipiperidinyl-1′-ylmethyl)pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.43-1.98 (m, 10H), 2.21 (m, 2H), 3.01 (m, 4H),3.43 (m, 3H), 4.12 (s, 2H), 5.16 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34(m, 6H), 9.85 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.7; ¹³C NMR (75MHz, DMSO) δ 21.6, 22.9, 23.8, 49.6, 50.5, 51.8, 53.0, 59.5, 108.0,127.8, 128.0, 128.2, 128.9, 137.3, 147.5, 157.6; ES MS (M+1) 382.4; HRMSCalcd. For C₂₃H₃₁N₃O₂, 383.51. Found (M+1) 382.25.

1-Benzyl-3-hydroxy-4-[(3,4-dihydroquinolin-1(2H)-yl)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 3.13 (t, J=6.3 Hz, 2H), 3.52 (m, 2H), 4.28 (s,2H), 4.41 (s, 2H), 5.18 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.23-7.41 (m,10H), 10.15 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.9; ¹³C NMR (75MHz, DMSO) δ 25.4; 49.3, 51.8, 52.7, 52.9, 107.6, 11.6, 116.8, 126.9,127.0, 127.9, 128.0, 128.1, 128.2, 128.8, 128.9, 131.7, 137.3, 147.3,157.6; ES MS (M+1) 347.40; HRMS Calcd. For C₂₂H₂₂N₂O₂, 346.42. Found(M+1) 347.17.

Methyl1-[(1-benzyl-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl)methyl]pyrrolidine-2-carboxylate

¹H NMR (300 MHz, DMSO) δ 2.01 (m, 3H), 2.45 (m, 1H), 3.26 (m, 1H), 3.53(m, 1H), 3.69 (s, 3H), 4.30 (m, 3H), 5.17 (s, 2H), 6.27 (d, 6.9 Hz, 1H),7.35 (m, 6H), ¹⁹F NMR (252 MHz, DMSO) δ 88.3; 13C NMR (75 MHz, DMSO) δ;ES MS (M+1) 343.20; HRMS Calcd. For C₁₉H₂₂N₂O₄, 342.39. Found (M+1)

1-Benzyl-3-hydroxy-4-{[2-(methoxymethyl)pyrrolidin-1-yl]methyl}pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.71 (m, 1H), 1.84 (m, 1H), 1.99 (m, 1H), 2.15(m, 1H), 3.19 (m, 1H), 3.30 (s, 3H), 3.41 (m, 1H), 3.62 (m, 2H), 3.77(m, 1H), 4.15 (m, 1H), 4.39 (m, 1H), 5.17 (s, 2H), 6.34 (d, J=7.2 Hz,1H), 7.34 (m, 6H); 9.60 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.3;¹³C NMR (75 MHz, DMSO) δ; ES MS (M+1) 329.2; HRMS Calcd. For C₁₉H₂₄N₂O₃,328.41. Found (M+1)

1-Benzyl-3-hydroxy-4-{[2-(pyrdin-2-yl)pyrrolidin-1-yl]methyl}pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 2.12 (m, 4H), 3.39 (m, 1H), 3.63 (m, 1H), 4.07(m, 2H), 4.60 (m, 1H), 5.10 (m, 2H), 6.15 (d, J=6.9 Hz, 1H), 7.33 (m,6H), 7.44 (m, 1H), 8.05 (d, J=8.1 Hz, 1H), 8.59 (d, J=4.8 Hz, 1H), 8.74(s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.0; ES MS (M+1) 362.22; HRMS Calcd.For C₂₂H₂₃N₃O₂, 361.44. Found (M+1).

1-Benzyl-3-hydroxy-4-[4-(6-chloropyridazin-3-yl)piperazin-1-ylmethyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 3.18 (m, 2H), 3.48 (m, 4H), 4.19 (s, 2H), 4.46(m, 2H), 5.16 (s, 2H), 6.62 (d, J=7.2 Hz, 1H), 7.35 (m, 6H), 7.48 (m,1H), 7.68 (m, 1H), 11.5 (broad s, 1H); ¹³C NMR (75 MHz, DMSO) δ 42.1,50.3, 51.9, 52.5, 108.2, 116.2; 118.0, 128.0, 128.2, 128.9, 129.8,137.3, 147.4, 157.6, 158.8; ES MS (M+1) 476.09. HRMS Calcd. ForC₂₁H₂₂ClN₅N₃O₂, 411.88. Found (M+1) 412.76.

1-Benzyl-3-hydroxy-4-[4-(2-methoxyphenyl)piperazin-1-ylmethyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 2.95 (m, 2H), 3.30 (m, 2H), 3.48 (m, 4H), 3.80(s, 3H), 4.25 (s, 2H), 5.18 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 6.93 (m,2H), 7.01 (m, 2H), 7.34 (m, 6H); ¹⁹F NMR (252 MHz, DMSO) δ 88.5; 13C NMR(75 MHz, DMSO) δ 47.2, 51.8, 53.0, 55.3, 108.1, 112.2, 114.8, 116.2,118.6, 121.2, 123.8, 127.8, 128.0, 128.9, 137.3, 139.6, 147.5, 152.2,157.6; ES MS (M+1) 405.82; HRMS Calcd. For C₂₄H₂₇N₃O₃, 405.49. Found(M+1) 406.21.

Category III of the disclosed prolyl hydroxylase inhibitors relates tocompounds having the formula:

R¹ and R² are each independently hydrogen or substituted orunsubstituted C₁-C₁₀ linear or branched alkyl, wherein the alkyl unitcan be substituted by one or more units independently chosen from:

i) C₁-C₈ linear, C₃-C₈ branched, or C₃-C₈ cyclic alkoxy;

ii) hydroxy;

iii) halogen;

iv) cyano;

v) amino, C₁-C₈ mono-alkylamino, C₁-C₈ di-alkylamino;

vi) —SR⁴⁰R⁴⁰ is hydrogen or C₁-C₄ linear or C₃-C₄ branched alkyl; vii)substituted or unsubstituted C₆ of C₁₀ aryl;

-   -   viii) substituted or unsubstituted C₁-C₉ heterocyclic; or    -   ix) substituted or unsubstituted C₁-C₉ heteroaryl.

Table III herein below provides non-limiting examples of compoundsencompassed by this category.

TABLE III No. R¹ R² C1 benzyl hydrogen C2 4-methoxybenzyl hydrogen C34-fluorobenzyl hydrogen C4 4-chlorobenzyl hydrogen C5 4-methylbenzylhydrogen C6 2-(pyridin-2-yl)ethyl hydrogen C7 [1,3]dioxolan-2-ylmethylhydrogen C8 tetrahydrofuran-2-ylmethyl hydrogen C9 2-methoxyethylhydrogen C10 1-hydroxy-2-methylpropan-2-yl hydrogen C11pyridin-4-ylmethyl hydrogen C12 furan-2-ylmethyl hydrogen C132-(methylthio)ethyl hydrogen C14 1-phenylethyl hydrogen C153-imidazol-1-ylpropyl hydrogen C16 cycloheptyl hydrogen C174-methylcyclohexyl hydrogen C18 1-benzylpiperidin-4-yl hydrogen C19azepan-2-on-3-yl hydrogen C20 1-benzylpyrrolidin-3-yl hydrogen C21benzyl methyl C22 4-methoxybenzyl methyl C23 4-fluorobenzyl methyl C244-chlorobenzyl methyl C25 4-methylbenzyl methyl C262-(pyridin-2-yl)ethyl methyl C27 [1,3]dioxolan-2-ylmethyl methyl C28tetrahydrofuran-2-ylmethyl methyl C29 2-methoxyethyl methyl C301-hydroxy-2-methylpropan-2-yl methyl C31 pyridin-4-ylmethyl methyl C32furan-2-ylmethyl methyl C33 2-(methylthio)ethyl methyl C34 1-phenylethylmethyl C35 3-(1H-imidazol-1-yl)propyl methyl C36 cycloheptyl methyl C374-methylcyclohexyl methyl C38 1-benzylpiperidin-4-yl methyl C39azepan-2-on-3-yl methyl C40 1-benzylpyrrolidin-3-yl methyl

The disclosed compounds of this category can be prepared by theprocedure outlined herein below in Scheme II and described in Example 2.

Reagents and conditions: (a)(i) HCHO, EtOH; 0.5 hr (ii)3-(1-H-imidazol-1-yl)propan-1-amine; 2 hr.

Example 21-Benzyl-3-hydroxy-4-{[3-(1-H-imidazol-1-yl)propylamino]methyl}-pyridin-2(1H)-one(6)

N-Benzyl-3-hydroxypyridin-2(1H)-one (5) can be prepared according toExample 1 by substituting benzyl bromide or benzyl chloride into step(b) for (4-chloro)benzyl chloride.

-   1-Benzyl-3-hydroxy-4-{[3-(1-H-imidazol-1-yl)propylamino]methyl}pyridin-2(1H)-one    (6): N-Benzyl-3-hydroxypyridin-2(1H)-one (5) (250 mg, 1.23 mmol) and    formaldehyde (200 mg, 273 eq.) are combined in aqueous ethanol (10    mL) and stirred for 30 minutes. 3-(1-H-Imidazol-1-yl)propan-1-amine    (340 mg, 2.7 mmol) is then added and the reaction stirred for 12    hours. The solvent is removed by evaporation and the residue    dissolved in methanol (2 mL) and purified via prep HPLC eluting with    water/acetonitrile to afford the desired product as the    trifluoroacetate salt. ¹H NMR (300 MHz, DMSO) δ 2.19 (m, 2H), 2.97    (m, 2H), 4.02 (s, 2H), 4.30 (t, J=6.6 Hz, 2H); 5.17 (s, 2H), 6.30    (d, J=6.9 Hz, 1H), 7.36 (m, 6H), 7.26 (s, 1H), 7.76 (s, 1H), 9.03    (s, 1H), 9.11 (s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.5; ¹³C NMR (75    MHz, DMSO) δ 26.5, 44.0, 46.0, 51.8, 106.8, 118.7, 120.5, 122.2,    127.9, 128.2, 128.9, 135.8, 137.4, 146.0, 158.2; ES MS (M+1) 339.05;    HRMS Calcd. For C₁₉H₂₂N₄O₂, 338.44. Found (M+1) 339.18.

The following are further non-limiting examples of this aspect of thedisclosed HIF-1α prolyl hydroxylase inhibitors.

1-Benzyl-3-hydroxy-4-(benzylaminomethyl)pyridin-2(1H)-one

¹HNMR (300 MHz, DMSO) δ 4.01 (s, 2H), 4.20 (s, 2H), 5.16 (s, 2H), 6.34(d, J=7.2 Hz, 1H), 7.36 (m, 11H), 9.16 (broad s, 1H); ¹⁹FNMR (252 MHz,DMSO) δ 88.6; ¹³C NMR (75 MHz, DMSO) 6; ES MS (M+1) 321.16; Anal. Calcd.For C₂₂H₂₁F₃N₂O₄, C, 60.83; H, 4.87; N, 6.45. Found: C, 60.75; H, 4.56;N, 6.34.

1-Benzyl-3-hydroxy-4-{[(2-(pyridin-2-yl)ethylamino]methyl}pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 3.26 (m, 2H), 3.37 (m, 2H), 4.08 (s, 2H), 5.17(s, 2H); 6.34 (d, J=7.2 Hz, 1H), 7.38 (m, 6H), 7.86 (d, J=5.7 Hz, 2H),8.84 (m, 2H), 9.32 (broad s, 1H); ¹⁹FNMR (252 MHz, DMSO) δ 88.6; ¹³C NMR(75 MHz, DMSO) δ 31.5, 44.1, 46.3, 51.8, 106.9, 114.8, 127.1, 128.1,128.8, 137.4, 143.8, 146.1, 155.3, 157.5, 158.4; ES MS (M+1) 336.18;HRMS Calcd. For C₂₀H₂₁N₃O₂, 335.40. Found: 336.16.

1-Benzyl-3-hydroxy-4-{[(tetrahydrofuran-2-ylmethyl)amino]methyl}pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.56 (m, 1H), 1.86 (m, 2H), 1.99 (m, 1H), 2.92(m, 1H), 3.05 (m, 1H), 3.80 (m, 2H), 4.09 (m, 3H), 5.16 (s, 2H), 6.34(d, J=7.2 Hz, 1H), 7.34 (m, 6H); 8.91 (broad s, 1H); ¹⁹F NMR (252 MHz,DMSO) δ 88.5; ¹³C NMR (75 MHz, DMSO) δ; ES MS (M+1) 315.16; HRMS. Calcd.For C₁₈H₂₂N₂O₃, 314.38. Found (M+1) 315.16.

1-Benzyl-3-hydroxy-4-[(2-methoxyethylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 3.13 (broad s, 2H), 3.30 (s, 3H), 3.59 (t,J=5.4 Hz, 2H), 4.02 (s, 2H), 5.16 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34(m, 6H), 8.91 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.4; ¹³C NMR(252 MHz, DMSO) δ; ES MS (M+1) 289.13; HRMS Calcd. For C₁₆H₂₀N₂O₃,288.34. Found (M+1) 289.15.

1-Benzyl-3-hydroxy-4-[(1-hydroxy-2-methylpropan-2-ylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.27 (s, 6H), 3.49 (s, 2H), 3.95 (s, 2H), 5.17(s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34 (m, 6H), 8.47 (broad s, 2H), 9.94(broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.7; 13C NMR (75 MHz, DMSO) δ;ES MS (M+1) 303.19; HRMS Calcd. For C₁₇H₂₂N₂O₃, 302.37. Found (M+1)303.17.

1-Benzyl-3-hydroxy-4-[(pyridin-4-ylmethylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 4.07 (s, 2H), 4.32 (s, 2H), 5, 16 (s, 2H), 6.34(d, J=7.2 Hz, 1H), 7.34 (m, 6H); 7.62 (d, J=5.7 Hz, 2H), 8.71 (d, J=4.5Hz, 2H); ¹⁹F NMR (252 MHz, DMSO) δ 88.0; ¹³C NMR (75 MHz, DMSO) δ; ES MS(M+1) 322.17; HRMS Calcd. For C₁₉H₁₉N₃O₂, 321.37. Found (M+1) 322.15.

1-Benzyl-3-hydroxy 4-{[(furan-2-ylmethyl)amino]methyl}pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 4.00 (s, 2H), 4.28 (s, 2H), 5.16 (s, 2H), 6.27(d, J=6.9 Hz, 1H), 6.54 (m, 1H), 6.65 (m, 1H), 7.34 (m, 6H), 7.80 (m,1H), 9.27 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.3; ¹³C NMR (75MHz, DMSO) δ; ES MS (M+1) 323.15; HRMS Calcd. For C₁₈H₁₈N₂O₃, 310.35.Found (M+1)

1-Benzyl-3-hydroxy-4-{[2-(methylthio)ethylamino]methyl}pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 2.10 (s, 3H), 2.74 (t, J=6.9 Hz, 2H), 3.16 (t,J=8.1 Hz, 2H), 4.05 (s, 2H), 5.17 (s, 2H), 6.34 (d, J=7.2 Hz, 1H), 7.34(m, 6H), ¹⁹F NMR (252 MHz, DMSO) δ 89.0; ES MS (M+1) 305.14, HRMS Calcd.For C₁₆H₂₀N₂O₂S, 304.41. Found (M+1)

1-Benzyl-3-hydroxy-4-[(4-methoxybenzylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 3.70 (s, 3H), 3.98 (s, 2H), 4.13 (s, 2H), 5.16(s, 2H), 6.28 (d, J=7.5 Hz, 1H), 7.00 (d, J=9.0 Hz, 4H), 7.34 (m, 6H);9.07 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 89.0; ES MS (M+1) 351.10;HRMS Calcd. For C₂₁H₂₂N₂O₃, 350.41. Found (M+1) 351.17.

1-Benzyl-3-hydroxy-4-[(1-phenylethylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.59 (d, J=7.2 Hz, 3H), 3.71-3.93 (m, 2H), 4.45(m, 1H), 5.15 (s, 2H), 6.28 (d, J=7.5 Hz, 1H), 7.34 (m, 11H); ¹⁹F NMR(252 MHz, DMSO) δ 88.9; ¹³C NMR (75 MHz, DMSO) δ 19.6, 42.5, 51.7, 58.0,106.8, 119.3, 128.0, 128.1, 128.2, 128.9, 129.3, 129.4, 137.3, 145.9,158.3; ES MS (M+1) 335.13; HRMS Calcd. For C₂₁H₂₂N₂O₂, 334.41. Found(M+1) 335.17.

1-Benzyl-3-hydroxy-4-(cycloheptylaminomethyl)pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.55 (m, 10H), 2.03 (m, 2H), 3.18 (s, 1H), 3.99(m, 2H), 5.17 (s, 2H), 6.32 (d, J=6.9 Hz, 1H), 7.35 (m, 6H), 8.65 (broads, 2H), 9.98 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.6; ¹³C NMR (75MHz, DMSO) δ 23.0, 27.2, 30.4, 41.6, 51.7, 58.9, 107.0, 111.7, 127.9,128.0, 128.2, 128.8, 137.4, 146.0, 157.5; ES MS (M+1) 327.13; HRMSCalcd. For C₂₀H₂₆N₂O₂, 326.43. Found (M+1) 327.20.

1-Benzyl-3-hydroxy-4-[(4-methylcyclohexylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 0.93 (d, J=6.9 Hz, 3H), 1.38 (m, 4H), 1.74 (m,4H), 2.05 (m, 1H), 3.10 (m, 1H), 4.01 (s, 2H), 5.17 (s, 2H), 6.31 (m,1H), 7.34 (m, 6H), 8.05 (broad s, 2H), 9.98 (broad s, 1H); ¹⁹F NMR (252MHz, DMSO) δ 88.9; ES MS (M+1) 327.14; HRMS Calcd. For C₂₀H₂₆N₂O₂,326.43. Found (M+1) 372.20.

1-Benzyl-3-hydroxy-4-[(1-benzylpiperidin-4-ylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.77 (m, 2H), 2.31 (m, 2H), 2.98 (m, 2H), 3.30(m, 3H), 3.46 (m, 2H), 4.03 (s, 2H), 0.29 (s, 2H), 5.16 (s, 2H), 6.30(d, J=7.5 Hz, 1H), 7.34 (m, 6H), 7.49 (s, 5H), 9.12 (broad s, 1H), 10.05(broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.8; ¹³C NMR (75 MHz, DMSO) δ27.1, 43.4, 51.8, 52.1, 54.2, 54.7, 57.6, 106.9, 118.5, 128.0, 128.1,128.8, 129.3, 129.8, 130.7, 131.3, 137.3, 146.2, 157.4; ES MS (M+1)404.56; HRMS Calcd. For C₂₅H₂₈N₃O₂, 403.52. Found (M+1) 404.23.

3-[(1-Benzyl-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl)methylamino]azepan-2-one

¹H NMR (300 MHz, DMSO) δ 1.25 (m, 1H), 1.59 (m, 2H), 1.74 (m, 1H), 1.92(m, 1H), 2.10 (m, 1H), 3.18 (m, 3H), 4.03 (s, 2H), 4.2 (m, 1H), 5.17 (s,2H), 6.33 (d, J=7.5 Hz, 1H), 7.34 (m, 6H), 8.31 (t, J=5.4 Hz, 1H), 9.07(broad s, 2H), 9.90 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.4; ¹³CNMR (75 MHz, DMSO) δ 27.0, 27.2, 28.4, 43.4, 51.7, 59.3, 107.1, 118.9,127.8, 127.9, 128.1, 128.9, 137.4, 146.0, 157.5, 166.3; ES MS (M+1)342.01; HRMS Calcd. For C₁₉H₂₃N₃O₃, 341.40. Found (M+1) 342.18.

1-Benzyl-3-hydroxy-4-[(1-benzylpyrrolidin-3-ylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 2.22 (m, 2H), 2.42 (m, 1H), 3.39 (m, 3H), 3.68(m, 1H), 4.06 (s, 2H), 4.39 (s, 2H), 5.17 (s, 2H), 6.33 (d, J=7.5 Hz,1H), 7.30-7.52 (m, 11H); ¹⁹F NMR (252 MHz, DMSO) δ 88.5; ¹³C NMR (75MHz, DMSO) δ 27.1, 43.4, 51.8, 52.1, 54.2, 54.7, 57.5, 106.9, 118.5,128.0, 128.8, 129.3, 129.8, 130.7, 131.3, 137.3, 146.2, 157.5; ES MS(M+1) 390.14; HRMS Calcd. For C₂₄H₂₇N₃O₂, 389.49. Found (M+1) 390.21.

(R)-1-Benzyl-3-hydroxy-4-[(1-phenylethylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.58 (d, J=6.9 Hz, 3H), 3.74 (m, 2H), 4.44 (m,1H), 5.14 (s, 2H), 6.23 (d, J=7.2 Hz, 1H), 7.35 (m, 6H); ¹⁹F NMR (252MHz, DMSO) δ 89.4; ¹³C NMR (75 MHz, DMSO) δ 19.6, 42.6, 51.7, 58.0,106.9, 18.7, 128.0, 128.1, 128.8, 129.3, 129.4, 137.2, 137.4, 145.9,157.5; ES MS (M+1) 335.13; Anal. Calcd. For C₂₁H₂₂N₂O₂, 334.41. Found(M+1) 335.31.

1-Benzyl-3-hydroxy-4-[([1,3]-dioxolan-2-ylmethylmethylamino)methyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 2.81 (s, 3H), 3.35 (d, J=3.9 Hz, 2H), 3.89 (m,2H), 4.01 (m, 2H), 4.21 (m, 2H), 5.17 (s, 2H); 5.27 (t, J=3.9 Hz, 1H),6.34 (d, J=7.2 Hz, 1H), 7.35 (m, 6H); ¹⁹F NMR (252 MHz, DMSO) δ 88.5;¹³C NMR (75 MHz, DMSO) δ; ES MS (M+1) 331.18; HRMS Calcd. ForC₁₈H₂₂N₂O₄, 330.38. Found (M+1) 331.16.

Category IV of the disclosed prolyl hydroxylase inhibitors relates tocompounds having the formula:

wherein A represents a ring optionally substituted by one or more R²⁰⁰units. Table IV provides non-limiting examples of this category.

TABLE IV No. A ring D1 pyrrolidin-1-yl D2 3-hydroxypyrrolidin-1-yl D32-(pyrdin-2-yl)pyrrolidin-1-yl D4 2-methylcarboxypyrrolidin-1-yl D52-(methoxymethyl)pyrrolidin-1-yl D6 thiazolidin-3-yl D7 1H-imidazol-1-ylD8 piperidin-1-yl D9 4-benzylpiperidin-1-yl D10 1,4′-bipiperidinyl-1′-ylD11 piperazin-1-yl D12 4-benzylpiperazin-1-yl D134-(2-methoxyphenyl)piperazin-1-ylmethyl D144-(6-chloropyridazin-3-yl)piperazin-1-yl D151,4-dioxa-8-azaspiro[4,5]dec-8-yl D16 morpholin-4-yl D17thiomorpholin-4-yl D18 azepan-1-yl D19 azocan-1-yl D203,4-dihydroquinolin-1(2H)-yl

The disclosed compounds of this category can be prepared by theprocedure outlined herein below in Scheme III and described in Example3.

Example 31-(4′-Methylbenzenesulfonyl)-3-hydroxy-4-(pyrrolidin-1-ylmethyl)pyridin-2(1H)-one(8) 1-(4′-Methylbenzenesulfonyl)-3-hydroxypyridin-2(1H)-one (7)

To stirred solution of 3-[(tert-butyldimethylsilyl)oxy]pyridin-2(1H)-one(1) (4.66 g, 20.7 mmol) in dry THF (150 mL), maintained at −78° C. undera dry nitrogen atmosphere is added n-butyl lithium (1.6 M solution inhexane, 21.0 mmol). After 20 minutes, 4-methyl-benzenesulfonyl chloride(3.95 g, 20.7 mmol) is added as a THF solution. The solution is allowedto warm to room temperature over one hour, the water (10 mL) is addedand the contents of the reaction vessel is extracted with EtOAc (3×),washed with brine (1×), dried over Na₂SO₄ and concentrated. The combinedorganic layers are dried over Na₂SO₄ and concentrated. The residue istaken up in ethanol (10 mL) and treated with conc. HCl (2 mL). Themixture is allowed to stir for 1 hour and the solvent is removed underreduced pressure to afford the desired compound as a white solid. ¹H NMR(300 MHz, DMSO) δ 2.43 (s, 3H), 6.14 (t, J=6.9 Hz, 1H), 6.76 (dd, J=7.65Hz, 1.5 Hz, 1H), 7.18 (dd, J=6.6 Hz, 1.8 Hz, 1H), 7.32 (d, J=7.3 Hz,2H), 7.98 (d, J=7.9 Hz, 2H).

1-(4′-Methylbenzenesulfonyl)-3-hydroxy-4-(pyrrolidin-1-ylmethyl)pyridin-2(1H)-one(8)

1-(4′-Methylbenzenesulfonyl)-3-hydroxypyridin-2(1H)-one (7) (250 mg,0.94 mmol) and formaldehyde (200 mg, 2.07 mmol) are combined in aqueousethanol (10 mL) and stirred for 30 minutes. Pyrrolidine (149 mg, 2.07mmol) is then added and the reaction stirred for 12 hours. The solventis removed by evaporation and the residue dissolved in methanol (5 mL)and purified via prep HPLC eluting with water/acetonitrile to afford thedesired product. ¹H NMR (300 MHz, DMSO) δ 1.87 (m, 2H), 1.99 (m, 2H),2.44 (s, 3H), 3.09 (m, 2H), 3.40 (m, 2H), 4.19 (s, 2H), 6.51 (d, J=7.5Hz, 1H), 7.51 (d, J=8.4 Hz, 1H), 7.76 (d, J=7.5 Hz, 1H), 7.98 (d, J=8.1Hz, 1H), 9.93 (broad s, 1H); ¹⁹F NMR (252 MHz, DMSO) δ 88.4; 13C NMR (75MHz, DMSO) δ 21.5, 22.7, 50.5, 53.7, 108.7, 118.6, 119.4, 128.4, 129.7,130.1, 133.1, 146.8, 147.7, 156.2; ES MS (M+1) 349.25; HRMS Calcd. ForC₁₇H₂₀N₂O₄S, 348.42. Found (M+1) 349.42.

The following are further non-limiting examples of prolyl hydroxylaseinhibitors according to this category.

1-(4′-Methylbenzenesulfonyl)-3-hydroxy-4-thiazolidin-3-ylmethylpyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 2.43 (s, 3H), 2.94 (t, J=6.6 MHz, 2H), 3.18 (t,J=6.0 Hz, 2H), 3.66 (s, 2H), 4.12 (s, 2H), 6.51 (d, J=7.5 Hz, 1H), 7.51(d, J=8.4 Hz, 1H), 7.76 (d, J=7.5 Hz, 1H), 7.98 (d, J=8.1 Hz, 1H), ¹⁹FNMR (252 MHz, DMSO) δ 87.9; ¹³C NMR (75 MHz, DMSO) δ 21.5, 21.9, 24.6,25.8, 50.3, 51.6, 108.7, 118.6, 120.8, 129.7, 130.1, 133.1, 146.9,148.1, 156.1, 158.4, 158.8; ES MS (M+1) 367.18; HRMS Calcd. ForC₁₆H₁₈N₂O₄S₂, 366.46. Found (M+1) 367.43.

1-(4′-Methylbenzenesulfonyl)-3-hydroxy-4-azocan-1ylmethylpyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.59 (m, 10H), 2.44 (s, 3H), 3.17 (m, 2H), 3.32(m, 2H), 4.15 (s, 2H), 6.51 (d, J=7.5 Hz, 1H), 7.51 (d, J=8.4 Hz, 1H),7.76 (d, J=7.5 Hz, 1H), 7.98 (d, J=8.1 Hz); ¹⁹F NMR (252 MHz, DMSO) δ88.7; ¹³C NMR (75 MHz, DMSO) δ 21.5, 21.9, 23.7, 24.6, 25.8, 50.3, 51.6,108.7, 118.9, 120.8, 129.8, 130.1, 133.1, 146.9, 148.2, 156.1; ES MS(M+1) 391.18; HRMS Calcd. For C₂₀H₂₆N₂O₄S, 390.18. Found (M+1) 391.23.

1-(4′Methylbenzenesulfonyl)-3-hydroxy-4-(4-phenylpiperazin-1-ylmethyl)-pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 2.43 (s, 3H), 3.13 (m, 8H), 3.43 (s, 2H), 6.47(d, J=7.5 Hz, 1H), 6.78 (t, J=7.2 Hz, 1H), 7.21 9 m, 2H), 7.50 (d, J=8.1Hz, 2H), 7.67 (d, J=7.8 Hz, 1H), 7.97 (d, J=8.4 Hz, 2H); ¹³C NMR (75MHz, DMSO) δ 21.5, 42.6, 45.6, 46.2, 50.8, 51.9, 109.6, 116.4, 116.8,117.7, 120.6, 121.1, 129.5, 129.6, 129.8, 130.1, 133.2, 146.8, 149.5,156.1; ES MS (M+1) 440.15; HRMS Calcd. For C₂₃H₂₅N₃O₅S, 439.53. Found(M+1) 440.16.

1-(4′-Methylbenzenesulfonyl)-3-hydroxy-4-[1,4′]Bipiperidinyl-1′-ylmethylpyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 1.43 (m, 1 h), 1.67 (m, 2H), 1.82 (m, 4H), 2.19(m, 2H), 2.44 (s, 3H), 2.94 (m, 4H), 3.39 (m, 2H), 3.54 (m, 3H), 4.06(s, 2H), 6.47 (d, J=8.1 Hz, 1H), 7.51 (d, J=8.1 Hz, 2H), 7.73 (d, 7.8Hz, 1H), 7.99 (d, J=8.4 Hz, 2H); ¹⁹F NMR (252 MHz, DMSO) δ 88.7; ¹³C NMR(75 MHz, DMSO) δ 21.4, 22.9, 23.6, 48.4, 49.5, 59.4, 109.3, 114.8,117.6, 120.5, 122.7, 129.7, 130.1, 133.1, 146.9, 148.6, 156.2; ES MS(M+1) 446.19; HRMS Calcd. For C₂₃H₃₁N₃O₄S, 445.58. Found (M+1) 446.21.

1-(4′-Methylbenzenesulfonyl)-3-hydroxy-4-[4-(6-chloropyridazin-3-yl)piperazin-1-ylmethyl]pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 2.44 (s, 3H), 3.17 (m, 2H), 3.46 (m, 4H), 4.17(s, 2H), 4.45 (m, 2H), 6.77 (d, J=7.8 Hz, 1H), 7.04 (m, 1H), 7.53 (m2H), 7.68 (m, 2H), 7.98 (m, 2H), 11.3 (broad s, 1H), ES MS (M+1) 476.92.HRMS Calcd. For C₂₁H₂₅ClN₅O₄S, 475.95. Found (M+1) 476.11.

Category V of HIF-1α prolyl hydroxylase inhibitors relates to compoundshaving the formula:

R represents from 1 to 5 optional substitutions for a phenyl ringhydrogen atom, R¹ and R² are each independently hydrogen or substitutedor unsubstituted C₁-C₁₀ linear or branched alkyl, wherein the alkyl unitcan be substituted by one or more units independently chosen from:

i) C₁-C₈ linear, C₃-C₈ branched, or C₃-C₈ cyclic alkoxy;

ii) hydroxy;

iii) halogen;

iv) cyano;

v) amino, C₁-C₈ mono-alkylamino, C₁-C₈ di-alkylamino;

vi) —SR⁴⁰R⁴⁰ is hydrogen or C₁-C₄ linear or C₃-C₄ branched alkyl;

vii) substituted or unsubstituted C₆ of C₁₀ aryl;

viii) substituted or unsubstituted C₁-C₉ heterocyclic; or

ix) substituted or unsubstituted C₁-C₉ heteroaryl.

Table V provides non-limiting examples of this category of HIF-1α prolylhydroxylase inhibitors.

TABLE V No. R R¹ R² E1 4-methyl benzyl hydrogen E2 4-methyl4-methoxybenzyl hydrogen E3 4-methyl 4-fluorobenzyl hydrogen E4 4-methyl4-chlorobenzyl hydrogen E5 4-methyl 4-methylbenzyl hydrogen E6 4-methyl2-(pyridin-2-yl)ethyl hydrogen E7 4-methyl [1,3]dioxolan-2-ylmethylhydrogen E8 4-methyl tetrahydrofuran-2-ylmethyl hydrogen E9 4-methyl2-methoxyethyl hydrogen E10 4-methyl 1-hydroxy-2-methylpropan-2-ylhydrogen E11 4-methyl pyridin-4-ylmethyl hydrogen E12 4-methylfuran-2-ylmethyl hydrogen E13 4-methyl 2-(methylthio)ethyl hydrogen E144-methyl 1-phenylethyl hydrogen E15 4-methyl 3-imidazol-1-ylpropylhydrogen E16 4-methyl cycloheptyl hydrogen E17 4-methyl4-methylcyclohexyl hydrogen E18 4-methyl 1-benzylpiperidin-4-yl hydrogenE19 4-methyl azepan-2-on-3-yl hydrogen E20 4-methyl1-benzylpyrrolidin-3-yl hydrogen E21 4-methyl benzyl methyl E22 4-methyl4-methoxybenzyl methyl E23 4-methyl 4-fluorobenzyl methyl E24 4-methyl4-chlorobenzyl methyl E25 4-methyl 4-methylbenzyl methyl E26 4-methyl2-(pyridin-2-yl)ethyl methyl E27 4-methyl [1,3]dioxolan-2-ylmethylmethyl E28 4-methyl tetrahydrofuran-2-ylmethyl methyl E29 4-methyl2-methoxyethyl methyl E30 4-methyl 1-hydroxy-2-methylpropan-2-yl methylE31 4-methyl pyridin-4-ylmethyl methyl E32 4-methyl furan-2-ylmethylmethyl E33 4-methyl carboxymethyl methyl E34 4-methyl2-(methylthio)ethyl methyl E35 4-methyl 1-phenylethyl methyl E364-methyl 3-imidazol-1-ylpropyl methyl E37 4-methyl cycloheptyl methylE38 4-methyl 4-methylcyclohexyl methyl E39 4-methyl1-benzylpiperidin-4-yl methyl E40 4-methyl azepan-2-on-3-yl methyl E414-methyl 1-benzylpyrrolidin-3-yl methyl

The disclosed compounds of this category can be prepared by theprocedure outlined herein below in Scheme IV and described in Examples4.

Example 41-(4′Methylbenzenesulfonyl)-3-hydroxy-4-[(benzylamino)methyl]-pyridin-2(1H)-one(9)1-(4′Methylbenzenesulfonyl)-3-hydroxy-4-(benzylaminomethyl)pyridin-2(1H)-one(9)

1-(4′-Methylbenzenesulfonyl)-3-hydroxypyridin-2(1H)-one (7) (250 mg,0.94 mmol) and formaldehyde (200 mg, 2.07 mmol) are combined in aqueousethanol (10 mL) and stirred for 30 minutes. Benzylamine (229 mg, 2.07mmol) is then added and the reaction stirred for 12 hours. The solventis removed by evaporation and the residue dissolved in methanol (5 mL)and purified via prep HPLC eluting with water/acetonitrile to afford thedesired product as the trifluoracetate salt. ¹H NMR (300 MHz, DMSO) d2.44 (s, 3H), 3.96 (s, 2H), 4.16 (s, 2H), 6.69 (d, J=8.1 Hz), 7.40 (m,7H), 7.52 (m, 1H), 7.73 (d, J=8.1 Hz, 1H), 7.97 (d, J=8.1 Hz, 1H), 9.71(broad s, 2H), 10.44 (broad s, 1H); ES MS (M+1) 396.67; HRMS Calcd. ForC₂₀H₂₀N₂O₄S, 384.45. Found (M+1) 385.12.

The following is a further non-limiting example of this category ofHIF-1α prolyl hydroxylase inhibitors.

1-(4′-Methylbenzenesulfonyl)-3-hydroxy-4-[(2-methoxyethylamino)methyl]-pyridin-2(1H)-one

¹H NMR (300 MHz, DMSO) δ 2.43 (s, 3H), 3.12 (m, 2H), 3.29 (s, 3H), 3.56(t, J=5.1 Hz, 2H), 3.99 (s, 2H), 6.51 (d, J=7.5 Hz, 1H), 7.51 (d, J=8.4Hz, 1H), 7.76 (d, J=7.5 Hz, 1H), 7.98 (d, J=8.1 Hz); ¹⁹F NMR (252 MHz,DMSO) δ 88.6; ¹³C NMR (75 MHz, DMSO) δ 21.5, 43.8, 46.2, 46.5, 58.5,67.2, 106.7, 119.2, 120.2, 123.9, 128.4, 129.7, 130.1, 133.1, 146.8,147.0, 156.0; ES MS (M+1) 353.12. HRMS Calcd. For C₁₆H₂₀N₂O₅S, 352.41.Found (M+1) 353.11.

Category VI of HIF-1α prolyl hydroxylase inhibitors relates to compoundshaving the formula:

wherein L is chosen from CH₂ or SO₂, and Z is substituted orunsubstituted phenyl. Non-limiting examples of inhibitors according tothis category are disclosed in Table VI below.

TABLE VI No. L Z F1 CH₂ 2-chlorophenyl F2 CH₂ 3-chlorophenyl F3 CH₂4-chlorophenyl F4 CH₂ 2-fluorophenyl F5 CH₂ 3-fluorophenyl F6 CH₂4-fluorophenyl F7 CH₂ 2,3-dichlorophenyl F8 CH₂ 2,4-dichlorophenyl F9CH₂ 2,5-dichlorophenyl F10 CH₂ 2,6-dichlorophenyl F11 CH₂3,4-dichlorophenyl F12 CH₂ 3,5-dichlorophenyl F13 CH₂ 2,3-difluorophenylF14 CH₂ 2,4-difluorophenyl F15 CH₂ 2,5-difluorophenyl F16 CH₂2,6-difluorophenyl F17 CH₂ 3,4-difluorophenyl F18 CH₂ 3,5-difluorophenylF19 CH₂ 2-cyanophenyl F20 CH₂ 3-cyanophenyl F21 CH₂ 4-cyanophenyl F22SO₂ 2-chlorophenyl F23 SO₂ 3-chlorophenyl F24 SO₂ 4-chlorophenyl F25 SO₂2-fluorophenyl F26 SO₂ 3-fluorophenyl F27 SO₂ 4-fluorophenyl F28 SO₂2,3-dichlorophenyl F29 SO₂ 2,4-dichlorophenyl F30 SO₂ 2,5-dichlorophenylF31 SO₂ 2,6-dichlorophenyl F32 SO₂ 3,4-dichlorophenyl F33 SO₂3,5-dichlorophenyl F34 SO₂ 2,3-difluorophenyl F35 SO₂ 2,4-difluorophenylF36 SO₂ 2,5-difluorophenyl F37 SO₂ 2,6-difluorophenyl F38 SO₂3,4-difluorophenyl F39 SO₂ 3,5-difluorophenyl F40 SO₂ 2-cyanophenyl F41SO₂ 3-cyanophenyl F42 SO₂ 4-cyanophenyl

The compounds encompassed within this category can be prepared accordingto Scheme I for Z equal to CH₂ and according to Scheme III for Z equalto SO₂.

Pharmacetically Acceptable Salts

The disclosed HIF-1α prolyl hydroxylase inhibitors can be in the form ofa pharmaceutically acceptable salt. Pharmaceutically acceptable saltscan be used by the formulator to provide a form of the disclosedinhibitor that is more compatible with the intended delivery of theinhibitor to a subject or for compatiblility of formulation.

The following are examples of procedures for preparing thepharmaceutically acceptable salt of the disclosed inhibitor,tert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate.

A suspension oftert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate(242 mg, 0.56 mmol) in MeOH (15 mL) was heated at reflux until ahomogeneous solution was obtained. Heating was stopped and 0.1N HCl (6.7mL, 1.2 eq.) was added while still hot and the solution was cooled toroom temperature. The volatiles were evaporated under reduced pressureand the amorphous residue was crystallized in acetone (5 mL). The solidwas collected by filtration.

A suspension oftert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate(217 mg, 0.5 mmol) in MeOH (15 mL) was heated at reflux untill ahomogeneous solution was obtained. Heating was stopped andmethanesulfonic acid (115.2 mg, 1.2 eq.) was added while still hot andthe solution was cooled to room temperature. The volatiles wereevaporated under reduced pressure and the amorphous residue wascrystallized in acetone (5 mL). The solid was collected by filtration.

Table VII herein below provides examples of pharmaceutically acceptablesalts oftert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylateformed from organic and inorganic acids. Start

TABLE VII Acid Yield Purity* M.P. (° C.) color Free base — 99.3% 183-184pink HCl 90% 99.7% 185-186 white H₂SO₄ 93% 99.7% 175 (dec.) slightlypink p-toluenesulfonyl 74% 99.8% 185-186 white methanesulfonyl 79% 99.9%155-157 white *HPLC analysis

¹H NMR analysis was used to determine the form of the salt, for example,that the mesylate salt formed herein above had the following formula:

¹H NMR analysis was used to determine at which site of the molecule saltformation was taking place. The chemical shifts for the protons on themethylene group bridging the piperazine and the pyridinone rings shiftedfrom 3.59 ppm in the free base to 4.31 ppm of the salt. In addition, thepiperazine methylene groups adjacent to the tertiary amine shifted from2.50 ppm to approximately 3.60 ppm. The chemical shifts for theremaining protons were largely unchanged. These data indicate that thetertiary amine nitrogen of the piperazine ring is protonated in saltforms. In addition, integration of the methyl protons of the methanesulfonyl unit relative to the core compound indicates the presence ofone equivalent of the acid.

The formulator can determine the solubility of the pharmaceuticallyacceptable salts of the disclosed inhibitors by any method desirable.The following is a non-limiting example of a procedure for evaluatingthe solubility of a salt of a disclosed inhibitor. A suspension oftert-butyl-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}-piperazine-1-carboxylatemethanesulfonate (26.6 mg) in distilled deionized water (3.0 mL) issonicated for 20 min with water bath temperature under 25° C. Thesuspension is filtered to remove any insoluble salt. The clear filtratesolution (200 μL) is diluted with distilled deionized water (800 μL) andsubjected to HPLC analysis. The following are results for thepharmaceutically acceptable salts outlined in Table VII above.

Solubility Salt (mg/mL) Purity* Free base ~0.001 99.3% hydrochloride 5.999.7% hydrogensulfonate 13.2 99.7% p-toluenesulfonate 2.3 99.8%methanesulfonate 16.6   99% *HPLC analysis

The following are non-limiting examples of other acids that can be usedto form pharmaceutically acceptable salts of the disclosed inhibitors:acetate, ctrate, maleate, succinate, lactate, glycolate and tartrate.

Further disclosed herein is a process for preparing the disclosed HIF-1αprolyl hydroxylase inhibitors, comprising:

-   -   a) protecting the hydroxyl moiety of hydroxypyridin-2(1H)-one to        prepare a protected pyridone having the formula:

-   -   -   wherein W represents a protecting group;

    -   b) reacting the protected pyridone with a compound having the        formula:

-   -   -   wherein R represents from 1 to 5 substitutions for hydrogen            as defined herein, the index n is an integer from 0 to 5, Q            is a leaving group, to form a O-protected N-benzyl pyridone            or N-sulfonylphenyl pyridone having the formula:

-   -   c) removing the protecting group from the O-protected N-benzyl        pyridone or N-sulfonylphenyl pyridone to form an N-benzyl        pyridone or N-sulfonylphenyl pyridone having the formula:

-   -   d) reacting an amine having the formula:

-   -   -   wherein R¹ and R² are the same as defined herein, with            formaldehyde to form an N-formylamine having the formula:

and

-   -   e) reacting the N-formylamine formed in step (d) with the        N-benzyl pyridone or N-sulfonylphenyl pyridone formed in        step (c) to form a compound having the formula:

Step (a) Preparation of an O-protected hydroxypyridin-2(1H)-one

Step (a) relates to the formation of an O-protectedhydroxypyridin-2(1H)-one having the formula:

W can be any protecting group. Non-limiting examples of protectinggroups include carbamates, for example, tert-butoxycarbonyl andmethoxycarbonyl, alkylsilanes, for example, trimethylsilyl andtert-butyldimethylsilyl, and the like.

Step (b) Preparation of O-protected N-benzyl hydroxypyridin-2(1H)-one orO-protected N-sulfonylphenyl hydroxypyridin-2(1H)-one

Step (b) relates to the formation of an O-protected N-benzylhydroxypyridin-2(1H)-one or O-protected N-sulfonylphenylhydroxypyridin-2(1H)-one having the formula

The protected hydroxypyridin-2(1H)-one formed in step (a) is reactedwith a compound having the formula:

wherein Q is a leaving group capable of being eliminated by theprotected hydroxypyridin-2(1H)-one ring nitrogen.

Step (c) Preparation of N-benzyl-3-hydroxypyridin-2(1H)-one orN-sulfonylphenyl-3-hydroxypyridin-2(1H)-one

Step (c) relates to the formation of anN-benzyl-3-hydroxypyridin-2(1H)-one orN-sulfonylphenyl-3-hydroxypyridin-2(1H)-one the having the formula:

Wherein the O-protected N-benzyl hydroxypyridin-2(1H)-one or O-protectedN-sulfonylphenyl hydroxypyridin-2(1H)-one formed in step (b) is reactedwith one or more reagents suitable for removing protecting group W in amanner compatible with any R substitutions for hydrogen on the phenylring.

Step (d) Preparation of an N-formylamine synthon

Step (d) relates to the formation of an N-formylamine synthon having theformula:

The N-formylamine is formed by reacting an amine having the formula:

with formaldehyde or a reagent capable of generating formaldehyde insitu.Step (e) Preparation of the disclosed HIF-1α prolyl hydroxylaseinhibitors

Step (e) relates to the formation of the final disclosed compoundshaving the formula:

by reacting the N-formylamine formed in step (d) with theN-benzyl-3-hydroxypyridin-2(1H)-one orN-sulfonylphenyl-3-hydroxypyridin-2(1H)-one formed in step (c).

FORMULATIONS Medicaments and Pharmaceutical Compositions

The present disclosure further relates to compositions or formulationsthat are useful for making a medicament or a pharmaceutical composition.The disclosed medicaments or pharmaceutical compositions comprising thedisclosed human protein HIF-1α prolyl hydroxylase inhibitors cancomprise:

-   -   a) an effective amount of one or more HIF-1α prolyl hydroxylase        inhibitors according to the present disclosure; and    -   b) one or more excipients.

Diseases or conditions affected by increased stabilization of HIF-1 byinhibition of HIF-1α prolyl hydroxylase include PVD, CAD, heart failure,ischemia, anemia, wound healing, antimicrobial activity, increasedphagocytosis, anti-cancer activity, and increase in the effectiveness ofvaccines.

For the purposes of the present disclosure the term “excipient” and“carrier” are used interchangeably throughout the description of thepresent disclosure and said terms are defined herein as, “ingredientswhich are used in the practice of formulating a safe and effectivepharmaceutical composition.”

The formulator will understand that excipients are used primarily toserve in delivering a safe, stable, and functional pharmaceutical,serving not only as part of the overall vehicle for delivery but also asa means for achieving effective absorption by the recipient of theactive ingredient. An excipient may fill a role as simple and direct asbeing an inert filler, or an excipient as used herein may be part of apH stabilizing system or coating to insure delivery of the ingredientssafely to the stomach. The formulator can also take advantage of thefact the compounds of the present disclosure have improved cellularpotency, pharmacokinetic properties, as well as improved oralbioavailability.

Non-limiting examples of compositions according to the presentdisclosure include:

-   -   a) from about 0.001 mg to about 1000 mg of one or more human        protein HIF-1α prolyl hydroxylase inhibitors according to the        present disclosure; and    -   b) one or more excipients.

Another example according to the present disclosure relates to thefollowing compositions:

-   -   a) from about 0.01 mg to about 100 mg of one or more human        protein prolyl HIF-1α prolyl hydroxylase inhibitors according to        the present disclosure; and    -   b) one or more excipients.

A further example according to the present disclosure relates to thefollowing compositions:

-   -   a) from about 0.1 mg to about 10 mg of one or more human protein        HIF-1αprolyl hydroxylase inhibitors according to the present        disclosure; and    -   b) one or more excipients.

A still further example of compositions according to the presentdisclosure comprise:

-   -   a) an effective amount of one or more human protein HIF-1α        prolyl hydroxylase inhibitors according to the present        disclosure; and    -   b) one or more chemotherapeutic agents or chemotherapeutic        compounds as further described herein.

A yet still further example of compositions according to the presentdisclosure comprise:

-   -   a) an effective amount of one or more human protein HIF-1α        prolyl hydroxylase inhibitors according to the present        disclosure; and    -   b) one or more vaccines for treatment of an infectious disease.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating anemia.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating increasing cellular immunity.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating cancer.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for increasing HIF-1 stabilization.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating anemia.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating peripheral vascular disease.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating wounds.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament that is an antimicrobial.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating atherosclerotic lesions.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating diabetes.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating hypertension.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating a disease affected by the level of vascularendothelial growth factor (VEGF), glyceraldehyde 3-phosphatedehydrogenase (GAPDH), and erythropoietin (EPO).

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating a disorder chosen from Crohn's disease andulcerative colitis, psoriasis, sarcoidosis, rheumatoid arthritis,hemangiomas, Osler-Weber-Rendu disease, or hereditary hemorrhagictelangiectasia, solid or blood borne tumors and acquired immunedeficiency syndrome.

The present disclosure further relates to the use of one or more of theHIF-1α prolyl hydroxylase inhibitors disclosed herein for making amedicament for treating a disorder chosen from diabetic retinopathy,macular degeneration, cancer, sickle cell anemia, sarcoid, syphilis,pseudoxanthoma elasticum, Paget's disease, vein occlusion, arteryocclusion, carotid obstructive disease, chronic uveitis/vitritis,mycobacterial infections, Lyme's disease, systemic lupus erythematosis,retinopathy of prematurity, Eales' disease, Behcet's disease, infectionscausing a retinitis or choroiditis, presumed ocular histoplasmosis,Best's disease, myopia, optic pits, Stargardt's disease, pars planitis,chronic retinal detachment, hyperviscosity syndrome, toxoplasmosis,trauma and post-laser complications, diseases associated with rubeosis,and proliferative vitreoretinopathy.

The disclosed compositions and the form of pharmaceutical preparationscomprising the HIF-1α prolyl hydroxylase inhibitors alone, or incombination with another drug or other therapeutic agent, inter alia,chemotherapeutic agent or chemotherapeutic compound, can vary accordingto the intended route of administration.

Orally administered preparations can be in the form of solids, liquids,emulsions, suspensions, or gels, or in dosage unit form, for example astablets or capsules. Tablets can be compounded in combination with otheringredients customarily used, such as tale, vegetable oils, polyols,gums, gelatin, starch, and other carriers. The HIF-1α prolyl hydroxylaseinhibitors can be dispersed in or combined with a suitable liquidcarrier in solutions, suspensions, or emulsions.

Parenteral compositions intended for injection, either subcutaneously,intramuscularly, or intravenously, can be prepared as liquids or solidforms for solution in liquid prior to injection, or as emulsions. Suchpreparations are sterile, and liquids to be injected intravenouslyshould be isotonic. Suitable excipients are, for example, water,dextrose, saline, and glycerol.

Administration of pharmaceutically acceptable salts of the substancesdescribed herein is included within the scope of the present disclosure.Such salts can be prepared from pharmaceutically acceptable non-toxicbases including organic bases and inorganic bases. Salts derived frominorganic bases include sodium, potassium, lithium, ammonium, calcium,magnesium, and the like. Salts derived from pharmaceutically acceptableorganic non-toxic bases include salts of primary, secondary, andtertiary amines, basic amino acids, and the like. For a helpfuldiscussion of pharmaceutical salts, see S. M. Berge et al., Journal ofPharmaceutical Sciences 66:1-19 (1977) the disclosure of which is herebyincorporated by reference.

Substances for injection can be prepared in unit dosage form in ampules,or in multidose containers. The HIF-1α prolyl hydroxylase inhibitors orcompositions comprising one or more HIF-1α prolyl hydroxylase inhibitorsto be delivered can be present in such forms as suspensions, solutions,or emulsions in oily or preferably aqueous vehicles. Alternatively, thesalt of the HIF-1α prolyl hydroxylase inhibitor can be in lyophilizedform for reconstitution, at the time of delivery, with a suitablevehicle, such as sterile pyrogen-free water. Both liquids as well aslyophilized forms that are to be reconstituted will comprise agents,preferably buffers, in amounts necessary to suitably adjust the pH ofthe injected solution. For any parenteral use, particularly if theformulation is to be administered intravenously, the total concentrationof solutes should be controlled to make the preparation isotonic,hypotonic, or weakly hypertonic. Nonionic materials, such as sugars, arepreferred for adjusting tonicity, and sucrose is particularly preferred.Any of these forms can further comprise suitable formulatory agents,such as starch or sugar, glycerol or saline. The compositions per unitdosage, whether liquid or solid, can contain from 0.1% to 99% ofpolynucleotide material.

METHODS Methods Relating to Stabilization of HIF-1

The eradication of invading microorganisms depends initially on innateimmune mechanisms that preexist in all individuals and act withinminutes of infection. Phagocytic cell types, including macrophages andneutrophils, play a key role in innate immunity because they canrecognize, ingest, and destroy many pathogens without the aid of anadaptive immune response. The effectiveness of myeloid cells in innatedefense reflects their capacity to function in low oxygen environments.Whereas in healthy tissues oxygen tension is generally 20-70 mm HG (i.e.2.5-9% oxygen), much lower levels (<1% oxygen) have been described inwounds and necrotic tissue foci (Arnold et al., Br J Exp Pathol 68, 569(1987); Vogelberg & Konig, Clin Investig 71, 466 (1993); Negus et al.,Am J Pathol 150, 1723 (1997)). It has also been shown (Zinkernagel A. S.et al., “Pharmacologic Augmentation of Hypoxia-Inducible Factor-1α withMimosine Boosts the Bactericidal Capacity of Phagocytes” J. InfectiousDiseases (2008):197: 214-217) that the HIF-1αagonist mimosine can boostthe capacity of human phagocytes and whole blood to kill the leadingpathogen Staphylococcus aureus in a dose-dependent fashion and reducethe lesion size in a murine model of S. aureus skin infection.

Macrophages are one population of effector cells involved in immuneresponses. Their role in natural immunity includes mediation ofphagocytosis, as well as release of cytokines and cytotoxic mediators.They also facilitate the development of acquired immunity throughantigen presentation and release of immunomodulatory cytokines. Althoughmacrophages are immune effectors, they are also susceptible to infectionby agents such as bacteria, protozoa, parasites, and viruses (TheMacrophage, C. E. Lewis & J. O'D. McGee. eds., IRL Press at OxfordUniversity Press, New York, N.Y., 1992). Viruses capable of infectingmacrophages include several RNA viruses such as measles virus (MV)(e.g., Joseph et al., J. Virol. 16, 1638-1649, 1975), respiratorysyncytial virus (RSV) (Midulla et al., Am. Rev. Respir. Dis. 140,771-777, 1989), and human immunodeficiency virus type 1 (HIV-1) (Meltzerand Gendelman, in Macrophage Biology and Activation, S. W. Russell andS. Gordon, eds., Springer-Verlag, New York, N.Y., pp. 239-263 (1992:Potts et al., Virology 175, 465-476, 1990).

Disclosed herein is a method for increasing HIF-1 stabilization in acell, comprising contacting a cell in vivo, in vitro, or ex vivo with aneffective amount of one or more of the disclosed HIF-1α prolylhydroxylase inhibitors.

Also disclosed herein are methods for increasing the cellular immuneresponse of a subject in need of increased cellular immunity, comprisingadministering to a subject in need with an effective amount of one ormore of the disclosed HIF-1α prolyl hydroxylase inhibitors.

Further disclosed herein are methods for increasing the cellular immuneresponse of a subject diagnosed with a medical condition causing adecreased cellular immunity, comprising administering to a subject inneed with an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

Yet further disclosed herein are methods for increasing the cellularimmune response of a subject diagnosed with a medical condition causinga decreased cellular immunity, comprising administering to a subject inneed with an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

Still further disclosed herein are methods for increasing the cellularimmune response of a subject having a medical condition causing adecreased cellular immunity, comprising administering to a subject inneed with an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

As such, the one or more HIF-1α prolyl hydroxylase inhibitor and anyco-administered compounds can be administered or contacted with a celltopically, buccally, orally, intradermally, subcutaneously, mucosally inthe eye, vagina, rectum, and nose, intravenously, and intramuscularly

Methods Relating to the Treatment of Cancer

As used herein cancer is defined herein as “an abnormal growth of cellswhich tend to proliferate in an uncontrolled way and, in some cases, tometastasize.” As such, both metastatic and non-metastatic cancers can betreated by the disclosed methods.

Disclosed are methods for treating cancer in a subject, comprisingadministering to a subject with a cancer with an effective amount of oneor more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

Also disclosed herein are methods for treating a subject diagnosed withcancer, co-administering to a subject one or more chemotherapeutic agentor chemotherapeutic compound together with one or more of the disclosedHIF-1α prolyl hydroxylase inhibitors.

The following are non-limiting examples of malignant and non-malignantcancers. Acute Lymphoblastic; Acute Myeloid Leukemia; AdrenocorticalCarcinoma; Adrenocortical Carcinoma, Childhood; Appendix Cancer; BasalCell Carcinoma; Bile Duct Cancer, Extrahepatic; Bladder Cancer; BoneCancer; Osteosarcoma and Malignant Fibrous Histiocytoma; Brain StemGlioma, Childhood; Brain Tumor, Adult; Brain Tumor, Brain Stem Glioma,Childhood; Brain Tumor, Central Nervous System AtypicalTeratoid/Rhabdoid Tumor, Childhood; Central Nervous System EmbryonalTumors; Cerebellar Astrocytoma; Cerebral Astrocytoma/Malignant Glioma;Craniopharyngioma; Ependymoblastoma; Ependymoma; Medulloblastoma;Medulloepithelioma; Pineal Parenchymal Tumors of IntermediateDifferentiation; Supratentorial Primitive Neuroectodermal Tumors andPineoblastoma; Visual Pathway and Hypothalamic Glioma; Brain and SpinalCord Tumors; Breast Cancer; Bronchial Tumors; Burkitt Lymphoma;Carcinoid Tumor; Carcinoid Tumor, Gastrointestinal; Central NervousSystem Atypical Teratoid/Rhabdoid Tumor; Central Nervous SystemEmbryonal Tumors; Central Nervous System Lymphoma; CerebellarAstrocytoma; Cerebral Astrocytoma/Malignant Glioma, Childhood; CervicalCancer; Chordoma, Childhood; Chronic Lymphocytic Leukemia; ChronicMyelogenous Leukemia; Chronic Myeloproliferative Disorders; ColonCancer; Colorectal Cancer; Craniopharyngioma; Cutaneous T-Cell Lymphoma;Esophageal Cancer; Ewing Family of Tumors; Extragonadal Germ Cell Tumor;Extrahepatic Bile Duct Cancer; Eye Cancer, Intraocular Melanoma; EyeCancer, Retinoblastoma; Gallbladder Cancer; Gastric (Stomach) Cancer;Gastrointestinal Carcinoid Tumor; Gastrointestinal Stromal Tumor (GIST);Germ Cell Tumor, Extracranial; Germ Cell Tumor, Extragonadal; Germ CellTumor, Ovarian; Gestational Trophoblastic Tumor; Glioma; Glioma,Childhood Brain Stem; Glioma, Childhood Cerebral Astrocytoma; Glioma,Childhood Visual Pathway and Hypothalamic; Hairy Cell Leukemia; Head andNeck Cancer; Hepatocellular (Liver) Cancer; Histiocytosis, LangerhansCell; Hodgkin Lymphoma; Hypopharyngeal Cancer; Hypothalamic and VisualPathway Glioma; Intraocular Melanoma; Islet Cell Tumors; Kidney (RenalCell) Cancer; Langerhans Cell Histiocytosis; Laryngeal Cancer; Leukemia,Acute Lymphoblastic; Leukemia, Acute Myeloid; Leukemia, ChronicLymphocytic; Leukemia, Chronic Myelogenous; Leukemia, Hairy Cell; Lipand Oral Cavity Cancer; Liver Cancer; Lung Cancer, Non-Small Cell; LungCancer, Small Cell; Lymphoma, AIDS-Related; Lymphoma, Burkitt; Lymphoma,Cutaneous T-Cell; Lymphoma, Hodgkin; Lymphoma, Non-Hodgkin; Lymphoma,Primary Central Nervous System; Macroglobulinemia, Waldenström;Malignant Fibrous Histiocytoma of Bone and Osteosarcoma;Medulloblastoma; Melanoma; Melanoma, Intraocular (Eye); Merkel CellCarcinoma; Mesothelioma; Metastatic Squamous Neck Cancer with OccultPrimary; Mouth Cancer; Multiple Endocrine Neoplasia Syndrome,(Childhood); Multiple Myeloma/Plasma Cell Neoplasm; Mycosis Fungoides;Myelodysplastic Syndromes; Myelodysplastic/Myelo-proliferative Diseases;Myelogenous Leukemia, Chronic; Myeloid Leukemia, Adult Acute; MyeloidLeukemia, Childhood Acute; Myeloma, Multiple; MyeloproliferativeDisorders, Chronic; Nasal Cavity and Paranasal Sinus Cancer;Nasopharyngeal Cancer; Neuroblastoma; Non-Small Cell Lung Cancer; OralCancer; Oral Cavity Cancer; Oropharyngeal Cancer; Osteosarcoma andMalignant Fibrous Histiocytoma of Bone; Ovarian Cancer; OvarianEpithelial Cancer; Ovarian Germ Cell Tumor; Ovarian Low MalignantPotential Tumor; Pancreatic Cancer; Pancreatic Cancer, Islet CellTumors; Papillomatosis; Parathyroid Cancer; Penile Cancer; PharyngealCancer; Pheochromocytoma; Pineal Parenchymal Tumors of IntermediateDifferentiation; Pineoblastoma and Supratentorial PrimitiveNeuroectodermal Tumors; Pituitary Tumor; Plasma Cell Neoplasm/MultipleMyeloma; Pleuropulmonary Blastoma; Primary Central Nervous SystemLymphoma; Prostate Cancer; Rectal Cancer; Renal Cell (Kidney) Cancer;Renal Pelvis and Ureter, Transitional Cell Cancer; Respiratory TractCarcinoma Involving the NUT Gene on Chromosome 15; Retinoblastoma;Rhabdomyosarcoma; Salivary Gland Cancer; Sarcoma, Ewing Family ofTumors; Sarcoma, Kaposi; Sarcoma, Soft Tissue; Sarcoma, Uterine; SézarySyndrome; Skin Cancer (Nonmelanoma); Skin Cancer (Melanoma); SkinCarcinoma, Merkel Cell; Small Cell Lung Cancer; Small Intestine Cancer;Soft Tissue Sarcoma; Squamous Cell Carcinoma, Squamous Neck Cancer withOccult Primary, Metastatic; Stomach (Gastric) Cancer; SupratentorialPrimitive Neuroectodermal Tumors; T-Cell Lymphoma, Cutaneous; TesticularCancer; Throat Cancer; Thymoma and Thymic Carcinoma; Thyroid Cancer;Transitional Cell Cancer of the Renal Pelvis and Ureter; TrophoblasticTumor, Gestational; Urethral Cancer; Uterine Cancer, Endometrial;Uterine Sarcoma; Vaginal Cancer; Vulvar Cancer; WaldenströmMacroglobulinemia; and Wilms Tumor

Further disclosed herein are methods for treating cancer in a subject,comprising co-administering to a subject, together with one or morechemotherapeutic agents or chemotherapeutic compounds, one or more ofthe disclosed HIF-1α prolyl hydroxylase inhibitors.

Also disclosed herein are methods for treating a subject diagnosed withcancer, co-administering to a subject, together with one or morechemotherapeutic agent or chemotherapeutic compound one or more of thedisclosed HIF-1α prolyl hydroxylase inhibitors.

A “chemotherapeutic agent” or “chemotherapeutic compound” is a chemicalcompound useful in the treatment of cancer. The chemotherapeutic canceragents that can be used in combination with the disclosed HIF-1αinhibitors include, but are not limited to, mitotic inhibitors (vincaalkaloids). These include vincristine, vinblastine, vindesine andNavelbine™ (vinorelbine, 5′-noranhydroblastine). In yet otherembodiments, chemotherapeutic cancer agents include topoisomerase Iinhibitors, such as camptothecin compounds. As used herein,“camptothecin compounds” include Camptosar™ (irinotecan HCl), Hycamtin™(topotecan HCl) and other compounds derived from camptothecin and itsanalogues. Another category of chemotherapeutic cancer agents that maybe used in the methods and compositions disclosed herein arepodophyllotoxin derivatives, such as etoposide, teniposide andmitopodozide. The present disclosure further encompasses otherchemotherapeutic cancer agents known as alkylating agents, whichalkylate the genetic material in tumor cells. These include withoutlimitation cisplatin, cyclophosphamide, nitrogen mustard, trimethylenethiophosphoramide, carmustine, busulfan, chlorambucil, belustine, uracilmustard, chlomaphazin, and dacarbazine. The disclosure encompassesantimetabolites as chemotherapeutic agents. Examples of these types ofagents include cytosine arabinoside, fluorouracil, methotrexate,mercaptopurine, azathioprime, and procarbazine. An additional categoryof chemotherapeutic cancer agents that may be used in the methods andcompositions disclosed herein include antibiotics. Examples includewithout limitation doxorubicin, bleomycin, dactinomycin, daunorubicin,mithramycin, mitomycin, mytomycin C, and daunomycin. There are numerousliposomal formulations commercially available for these compounds. Thepresent disclosure further encompasses other chemotherapeutic canceragents including without limitation anti-tumor antibodies, dacarbazine,azacytidine, amsacrine, melphalan, ifosfamide and mitoxantrone.

The disclosed HIF-1α prolyl hydroxylase inhibitors herein can beadministered in combination with other anti-tumor agents, includingcytotoxic/antineoplastic agents and anti-angiogenic agents.Cytotoxic/anti-neoplastic agents are defined as agents which attack andkill cancer cells. Some cytotoxic/anti-neoplastic agents are alkylatingagents, which alkylate the genetic material in tumor cells, e.g.,cis-platin, cyclophosphamide, nitrogen mustard, trimethylenethiophosphoramide, carmustine, busulfan, chlorambucil, belustine, uracilmustard, chlomaphazin, and dacabazine. Other cytotoxic/anti-neoplasticagents are antimetabolites for tumor cells, e.g., cytosine arabinoside,fluorouracil, methotrexate, mercaptopuirine, azathioprime, andprocarbazine. Other cytotoxic/anti-neoplastic agents are antibiotics,e.g., doxorubicin, bleomycin, dactinomycin, daunorubicin, mithramycin,mitomycin, mytomycin C, and daunomycin. There are numerous liposomalformulations commercially available for these compounds. Still othercytotoxic/anti-neoplastic agents are mitotic inhibitors (vincaalkaloids). These include vincristine, vinblastine and etoposide.Miscellaneous cytotoxic/anti-neoplastic agents include taxol and itsderivatives, L-asparaginase, anti-tumor antibodies, dacarbazine,azacytidine, amsacrine, melphalan, VM-26, ifosfamide, mitoxantrone, andvindesine.

Anti-angiogenic agents are well known to those of skill in the art.Suitable anti-angiogenic agents for use in the disclosed methods andcompositions include anti-VEGF antibodies, including humanized andchimeric antibodies, anti-VEGF aptamers and antisense oligonucleotides.Other known inhibitors of angiogenesis include angiostatin, endostatin,interferons, interleukin 1 (including α and β) interleukin 12, retinoicacid, and tissue inhibitors of metalloproteinase-1 and -2. (TIMP-1 and-2). Small molecules, including topoisomerases such as razoxane, atopoisomerase II inhibitor with anti-angiogenic activity, can also beused.

Other anti-cancer agents that can be used in combination with thedisclosed HIF-1αinhibitors include, but are not limited to: acivicin;aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin;altretamine; ambomycin; ametantrone acetate; aminoglutethimide;amsacrine; anastrozole; anthramycin; asparaginase; asperlin;azacitidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide;bisantrene hydrochloride; bisnafide dimesylate; bizelesin; bleomycinsulfate; brequinar sodium; bropirimine; busulfan; cactinomycin;calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicinhydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin;cisplatin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine;dacarbazine; dactinomycin; daunorubicin hydrochloride; decitabine;dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; docetaxel;doxorubicin; doxorubicin hydrochloride; droloxifene; droloxifenecitrate; dromostanolone propionate; duazomycin; edatrexate; eflornithinehydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine;epirubicin hydrochloride; erbulozole; esorubicin hydrochloride;estramustine; estramustine phosphate sodium; etanidazole; etoposide;etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine;fenretinide; floxuridine; fludarabine phosphate; fluorouracil;fluorocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabinehydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide;ilmofosine; interleukin II (including recombinant interleukin II, orrIL2), interferon alfa-2a; interferon alfa-2b; interferon alfa-n1;interferon alfa-n3; interferon beta-I a; interferon gamma-I b;iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole;leuprolide acetate; liarozole hydrochloride; lometrexol sodium;lomustine; losoxantrone hydrochloride; masoprocol; maytansine;mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate;melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium;metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin;mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride;mycophenolic acid; nocodazole; nogalamycin; ormaplatin; oxisuran;paclitaxel; pegaspargase; peliomycin; pentamustine; peplomycin sulfate;perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride;plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine;procarbazine hydrochloride; puromycin; puromycin hydrochloride;pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride;semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermaniumhydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin;sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantronehydrochloride; temoporfin; teniposide; teroxirone; testolactone;thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifenecitrate; trestolone acetate; triciribine phosphate; trimetrexate;trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracilmustard; uredepa; vapreotide; verteporfin; vinblastine sulfate;vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate;vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate;vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin;zinostatin; zorubicin hydrochloride. Other anti-cancer drugs include,but are not limited to: 20-epi-1,25 dihydroxyvitamin D3;5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol;adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine;amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine;anagrelide; anastrozole; andrographolide; angiogenesis inhibitors;antagonist D; antagonist G; antarelix; anti-dorsalizing morphogeneticprotein-1; antiandrogen, prostatic carcinoma; antiestrogen;antineoplaston; antisense oligonucleotides; aphidicolin glycinate;apoptosis gene modulators; apoptosis regulators; apurinic acid;ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane;atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron;azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat;BCR/ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactamderivatives; beta-alethine; betaclamycin B; betulinic acid; bFGFinhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide;bistratene A; bizelesin; breflate; bropirimine; budotitane; buthioninesulfoximine; calcipotriol; calphostin C; camptothecin derivatives;canarypox IL-2; capecitabine; carboxamide-amino-triazole;carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor;carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropinB; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost;cis-porphyrin; cladribine; clomifene analogues; clotrimazole;collismycin A; collismycin B; combretastatin A4; combretastatinanalogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8;cryptophycin A derivatives; curacin A; cyclopentanthraquinones;cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor;cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin;dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone;didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine;dihydrotaxol, 9-; dioxamycin; diphenyl spiromustine; docetaxel;docosanol; dolasetron; doxifluridine; droloxifene; dronabinol;duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab;eflornithine; elemene; emitefur; epirubicin; epristeride; estramustineanalogue; estrogen agonists; estrogen antagonists; etanidazole;etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide;filgrastim; finasteride; flavopiridol; flezelastine; fluasterone;fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane;fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate;galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathioneinhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin;ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine;ilomastat; imidazoacridones; imiquimod; immunostimulant peptides;insulin-like growth factor-1 receptor inhibitor; interferon agonists;interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-;iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RH retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen binding protein; sizofiran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatinstimalamer. In one embodiment, the anti-cancer drug is 5-fluorouracil,taxol, or leucovorin.

Methods Related to Treatment of Conditions Involving Microorganisms

Disclosed is a method for prophylactically treating a human or a mammalagainst infection by a microorganism, comprising administering to asubject an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

Further disclosed is a method for decreasing the virulence of amicroorganism when a human or a mammal is infected with a microorganism,comprising administering to a subject an effective amount of one or moreof the disclosed HIF-1α prolyl hydroxylase inhibitors.

Yet further disclosed is a method for treating an infection in a subjectcaused by a microorganism, comprising administering to a subject aneffective amount of one or more of the disclosed HIF-1α prolylhydroxylase inhibitors.

Still further disclosed is a method for treating a subject diagnosedwith an infection caused by a microorganism, comprising administering toa subject an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

Also disclosed is a method for preventing transmission of a diseasecaused by a microorganism from a subject to a subject, comprisingadministering to a subject an effective amount of one or more of thedisclosed HIF-1α prolyl hydroxylase inhibitors.

Still yet further disclosed is a method for preventing infection of ahuman or a mammal during a surgical procedure, comprising administeringto a subject an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

The microorganism can be any benign or virulent microorganism, forexample, bacteria, viruses, yeasts, fungi, or parasites. The followingare non-limiting examples of microorganisms that can be affected by thedisclosed HIF-1α prolyl hydroxylase inhibitors. By the term “affected”is meant, the virulence of the microorganism is reduced, diminished oreliminated. The cause of the reduction, diminishment, or elimination ofthe virulence can be from the stabilization of HIF-1 and/or from theincreased level of phagocytosis due to the administration of one or moreof the disclosed HIF-1α prolyl hydroxylase inhibitors.

Acinetobacter calcoaceticus, Acinetobacter haemolyticus, Aeromonashydrophilia, Agrobacterium tumefaciens, Bacillus anthracis, Bacillushalodurans, Bacillus subtilis, Bacteroides distasonis, Bacteroideseggerthii, Bacteroides fragilis, Bacteroides ovalus, Bacteroides 3452Ahomology group, Bacteroides splanchnicus, Bacteroides thetaiotaomicron,Bacteroides uniformis, Bacteroides vulgatus, Bordetella bronchiseptica,Bordetella parapertussis, Bordetella pertussis, Borrelia burgdorferi,Branhamella catarrhalis, Brucella melitensis, Burkholderia cepacia,Burkholderia pseudomallei, Campylobacter coli, Campylobacterfetus,Campylobacter jejuni, Caulobacter crescentus, Citrobacter freundii,Clostridium difficile, Clostridium perfringens, Corynebacteriumdiphtheriae, Corynebacterium glutamicum, Corynebacterium ulcerans,Edwardsiella tarda, Enterobacter aerogenes, Erwinia chrysanthemi,Enterobacter cloacae, Enterococcus faecalis, Enterococcus faecium,Escherichia coli, Francisella tularensis, Gardnerella vaginalis,Haemophilus ducreyi, Haemophilus haemolyticus, Haemophilus influenzae,Haemophilus parahaemolyticus, Haemophilus parainfluenzae, Helicobacterpylori, Klebsiella oxytoca, Klebsiella pneumoniae, Kluyveracryocrescens, Legionella pneumophila, Listeria innocua, Listeriamonocytogenes, Listeria welshimeri, Methanosarcina acetivorans,Methanosarcina mazei, Morganella morganii, Mycobacterium avium,Mycobacterium intracellulare, Mycobacterium leprae, Mycobacteriumtuberculosis, Mesorhizobium loti, Neisseria gonorrhoeae, Neisseriameningitidis, Pasteurella haemolytica, Pasteurella multocida,Providencia alcalifaciens, Providencia rettgeri, Providencia stuartii,Proteus mirabilis, Proteus vulgaris, Pseudomonas acidovorans,Pseudomonas aeruginosa, Pseudomonas alcaligenes, Pseudomonasfluorescens,Pseudomonas putida, Ralstonia solanacearum, Salmonella enterica subsp.enteridtidis, Salmonella enterica subsp. paratyphi, Salmonella enterica,subsp. typhimurium, Salmonella enterica, subsp. typhi, Serratiamarcescens, Shigella dysenteriae, Shigella flexneri, Shigella sonnei,Sinorhizobium meliloti, Staphylococcus aureus, Streptococcus criceti,Staphylococcus epidemmidis, Staphylococcus haemolyticus, Staphylococcushominis, Staphylococcus hyicus, Staphylococcus intermedius,Stenotrophomonas maltophilia, Staphylococcus saccharolyticus,Staphylococcus saprophyticus, Staphylococcus sciuri, Streptomycesavermitilis, Streptomyces coelicolor, Streptococcus agalactiae,Streptococcus pneumoniae, Streptococcus pyogenes Sulfobalblobussoffiataricus, Thermotoga maritima, Vibrio cholerae, Vibrioparahaemolyticus, Vogesella indigofera, Xanthomonas axonopodis,Xanthomonas campestris, Yersinia enterocolitica, Yersinia intermedia,Yersinia pestis, and Yersinia pseudotuberculosis

Methods Relating to Vaccination or Innoculation

Disclosed herein are methods for enhancing the effectiveness of avaccine, comprising co-administering to a subject a vaccine incombination with one or more HIF-1α prolyl hydroxylase inhibitors.

Non-limiting examples of vaccines are those for stimulating antibodiesagainst hepatitis, influenza, measles, rubella, tetanus, polio, rabies,and the like.

Therefore, the disclosed methods includes administering, or in the caseof contacting cells in vitro, in vivo or ex vivo, the one or more HIF-1αprolyl hydroxylase inhibitors and any co-administered compoundstopically, buccally, orally, intradermally, subcutaneously, mucosally inthe eye, vagina, rectum, and nose, intravenously, and intramuscularly.

The following are non-limiting examples of methods according to thepresent disclosure.

A method for increasing HIF-1 stabilization in a cell, comprisingcontacting a cell in vivo, in vitro, or ex vivo with an effective amountof one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

A method for increasing the cellular immune response of a subject inneed of increased cellular immunity, comprising administering to asubject in need with an effective amount of one or more of the disclosedHIF-1α prolyl hydroxylase inhibitors.

A method for increasing the cellular immune response of a subjectdiagnosed with a medical condition causing a decreased cellularimmunity, comprising administering to a subject in need an effectiveamount of one or more of the disclosed HIF-1α prolyl hydroxylaseinhibitors.

A method for increasing the cellular immune response of a subjectdiagnosed with a medical condition causing a decreased cellularimmunity, comprising administering to the subject an effective amount ofone or more of the disclosed HIF-1α inhibitors.

A method for increasing the cellular immune response of a subject havinga medical condition causing a decreased cellular immunity, said methodcomprising administering to the subject in need an effective amount ofone or more of the disclosed HIF-1α inhibitors.

A method for treating cancer in a subject, comprising administering tothe subject having cancer an effective amount of one or more of thedisclosed HIF-1α inhibitors.

A method for treating a subject diagnosed with cancer by stabilizing thelevel of cellular HIF-1 thereby increasing the immune response in thesubject, comprising administering to the subject diagnosed with cancerwith an effective amount of one or more HIF-1α inhibitors.

A method for treating a subject diagnosed with cancer by stabilizing thelevel of cellular HIF-1 thereby increasing the immune response in thesubject, comprising administering to the subject diagnosed with canceran effective amount of a composition comprising one or morechemotherapeutic agents or chemotherapeutic compounds and one or moreHIF-1α prolyl hydroxylase inhibitors.

A method for treating cancer in a subject by stabilizing the level ofcellular HIF-1 thereby increasing the immune response in the subject,comprising administering to the subject with cancer with an effectiveamount of one or more HIF-1α inhibitors.

A method for treating cancer in a subject by stabilizing the level ofcellular HIF-1 thereby increasing the immune response in the subject,said method comprising administering to the subject an effective amountof a composition comprising one or more chemotherapeutic agents orchemotherapeutic compounds and one or more HIF-1α prolyl hydroxylaseinhibitors.

A method for treating a subject diagnosed with cancer, comprisingadministering to the subject diagnosed with cancer with an effectiveamount of one or more HIF-1αinhibitors.

A method for treating a subject diagnosed with cancer, comprisingadministering to the subject diagnosed with cancer with an effectiveamount of a composition comprising one or more chemotherapeutic agentsor chemotherapeutic compounds and one or more HIF-1α prolyl hydroxylaseinhibitors.

A method for treating a subject diagnosed with cancer byco-administering to the subject one or more chemotherapeutic agents orchemotherapeutic compounds and one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

A method for treating a subject having cancer by co-administering to thesubject one or more chemotherapeutic agents or chemotherapeuticcompounds and one or more of the disclosed HIF-1α prolyl hydroxylaseinhibitors.

A method for prophylactically treating infection in a subject against aninfection, comprising administering to the subject an effective amountof one or more of the disclosed HIF-1α inhibitors.

A method for treating an infection in a subject caused by amicroorganism, comprising administering to the subject an effectiveamount of one or more of the disclosed HIF-1α prolyl hydroxylaseinhibitors.

A method for treating infection in a subject wherein the infection iscaused by a pathogen chosen from bacteria, viruses, yeasts, fungi, orparasites, comprising administering to the subject with an infectionwith an effective amount of one or more HIF-1α inhibitors.

A method for treating a subject diagnosed with an infection caused by amicroorganism, comprising administering to the subject an effectiveamount of one or more of the disclosed HIF-1α prolyl hydroxylaseinhibitors.

A method for preventing transmission of a disease caused by amicroorganism from one subject to another subject, comprisingadministering to the subject an effective amount of one or more of thedisclosed HIF-1α prolyl hydroxylase inhibitors.

A method for enhancing the effectiveness of a vaccine, comprisingco-administering to a subject a vaccine in combination with one or moreof the disclosed HIF-1α prolyl hydroxylase inhibitors.

A method for enhancing the effectiveness of a vaccine, comprisingadministering to a subject a composition comprising a vaccine incombination with one or more of the disclosed HIF-1α prolyl hydroxylaseinhibitors.

A method for inhibiting hypoxia-inducible factor-1alpha (HIF-1α)4-prolyl hydroxylase activity in a subject thereby stabilizing thecellular level of HIF-1, comprising administering to the subject aneffective amount of one or more of the disclosed HIF-1α 4-prolylhydroxylase inhibitors.

A method for modulating the cellular level of hypoxia-inducible factor-1(HIF-1) in a subject, comprising contacting the subject with aneffective amount of one or more an effective amount of one or more ofthe disclosed HIF-1α prolyl hydroxylase inhibitors.

A method for increasing the cellular level of HIF-1 in a subject duringa state of normoxia, comprising administering to a subject an effectiveamount of one or more an effective amount of one or more of thedisclosed HIF-1α prolyl hydroxylase inhibitors.

A method for treating anemia in a subject, comprising administering tothe subject an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

A method for treating a subject diagnosed as having anemia, comprisingadministering to the subject an effective amount of one or more of thedisclosed HIF-1αprolyl hydroxylase inhibitors.

A method for increasing angiogenesis in a subject, comprisingadministering to the subject an effective amount of one or more of thedisclosed HIF-1α prolyl hydroxylase inhibitors.

A method for treating a subject having a need for increasedangiogenesis, comprising administering to the subject an effectiveamount of one or more of the disclosed HIF-1α prolyl hydroxylaseinhibitors.

A method for treating sepsis in a subject, comprising administering tothe subject an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

A method for treating a subject diagnosed as having sepsis, comprisingadministering to a subject an effective amount of one or more HIF-1αprolyl hydroxylase inhibitors.

A method for treating peripheral vascular disease in a subject,comprising administering to a subject an effective amount of one or moreof the disclosed HIF-1αprolyl hydroxylase inhibitors.

A method for treating a subject diagnosed as having peripheral vasculardisease, comprising administering to a subject an effective amount ofone or more of the disclosed HIF-1α prolyl hydroxylase inhibitors.

A method for treating a wound in a subject, comprising administering toa subject an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

A method for treating a subject having a wound, comprising administeringto a subject an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

A method for preventing infection of a wound in a subject, comprisingadministering to a subject an effective amount of one or more of thedisclosed HIF-1α prolyl hydroxylase inhibitors.

A method for treating diabetes in a subject, comprising administering tothe subject an effective amount of one or more of the disclosed HIF-1αprolyl hydroxylase inhibitors.

A method for treating a subject diagnosed as having diabetes, comprisingadministering to the subject an effective amount of one or more of thedisclosed HIF-1αprolyl hydroxylase inhibitors.

A method for treating hypertension in a subject, comprisingadministering to the subject an effective amount of one or more of thedisclosed HIF-1α prolyl hydroxylase inhibitors.

A method for treating a subject diagnosed as having hypertension,comprising administering to the subject an effective amount of one ormore of the disclosed HIF-1αprolyl hydroxylase inhibitors.

PROCEDURES EGLN-1 Activity Assay

The EGLN-1 (or EGLN-3) enzyme activity is determined using massspectrometry (matrix-assisted laser desorption ionization,time-of-flight MS, MALDI-TOF MS. Recombinant human EGLN-1-179/426 isprepared as described above and in the Supplemental Data. Full-lengthrecombinant human EGLN-3 is prepared in a similar way, however it isnecessary to use the His-MBP-TVMV-EGLN-3 fusion for the assay due to theinstability of the cleaved protein. For both enzymes, the HIF-1α peptidecorresponding to residues 556-574 is used as substrate. The reaction isconducted in a total volume of 50 μL containing TrisCl (5 mM, pH 7.5),ascorbate (120 μM), 2-oxoglutarate (3.2 μM), HIF-1α(8.6 μM), and bovineserum albumin (0.01%). The enzyme, quantity predetermined to hydroxylate20% of substrate in 20 minutes, is added to start the reaction. Whereinhibitors are used, compounds are prepared in dimethyl sulfoxide at10-fold final assay concentration. After 20 minutes at room temperature,the reaction is stopped by transferring 10 μL of reaction mixture to 50μL of a mass spectrometry matrix solution (α-cyano-4-hydroxycinnamicacid, 5 mg/mL in 50% acetonitrile/0.1% TFA, 5 mM NH₄PO₄). Twomicroliters of the mixture is spotted onto a MALDI-TOF MS target platefor analysis with an Applied Biosystems (Foster City, Calif.) 4700Proteomics Analyzer MALDI-TOF MS equipped with a Nd:YAG laser (355 nm, 3ns pulse width, 200 Hz repetition rate). Hydroxylated peptide product isidentified from substrate by the gain of 16 Da. Data defined as percentconversion of substrate to product is analyzed in GraphPad Prism 4 tocalculate IC₅₀ values.

VEGF ELISA Assay

HEK293 cells are seeded in 96-well poly-lysine coated plates at 20,000cells per well in DMEM (10% FBS, 1% NEAA, 0.1% glutamine). Followingovernight incubation, the cells are washed with 100 μL of Opti-MEM(Gibco, Carlsbad, Calif.) to remove serum. Compound in DMSO is seriallydiluted (beginning with 100 μM) in Opti-MEM and added to the cells. Theconditioned media is analyzed for VEGF with a Quantikine human VEGFimmunoassay kit (R&D Systems, Minneapolis, Minn.). Optical densitymeasurements at 450 nm are recorded using the Spectra Max 250 (MolecularDevices, Sunnyvale, Calif.). Data defined as % of DFO stimulation isused to calculate EC₅₀ values with GraphPad Prism 4 software (San Diego,Calif.).

Mouse Ischemic Hindlimb Study

All animal work is conducted in accordance with the Guide for the Careand Use of Laboratory Animals (National Academy of Sciences; Copyright©1996). Used in these experiments were 9-10 week old male C57B1/6 micefrom Charles River Laboratory (Portage, Mich.). The mice are orallydosed with vehicle (aqueous carbonate buffer, 50 mM; pH 9.0) or with thecompound to be tested in vehicle at 50 mg/kg or 100 mg/kg. The animalsare dosed three times: day 1 at 8 AM and 5 PM, and on day 2 at 8 AM. Onehour after the first dose, unilateral arterial ligation is performedunder anesthesia using isoflurane. The femoral artery is ligatedproximal to the origin of the popliteal artery. The contralateral limbundergoes a sham surgical procedure. Ligation is performed in analternating fashion between right and left hindlimbs. Two hours afterthe 8 AM dosing on day 2, blood is obtained by ventricular stick whilethe mice are anesthetized with isoflurane. Serum samples for EPOanalysis are obtained using gel clot serum separation tubes. Heart,liver, and gastrocnemius muscles are harvested, snap-frozen in liquidnitrogen, and stored in −80° C. until use.

Mouse Serum EPO Assay

The mouse serum EPO is detected using Mouse Quantikine ErythropoietinELISA kit from R&D Systems according to manufacturer's instructions.

Mouse Tissue HIF Western Blot Analysis

Tissues from mice stored at −80° C. are powdered with mortar and pestlechilled with liquid nitrogen. Nuclear extracts are prepared using anNE-PER kit (Pierce Biotechnology). For immunoprecipitation, nuclearextract is added to monoclonal antibody to HIF-1α(Novus, Littleton,Colo.) at a tissue to antibody ratio of 200:1. The suspension isincubated in a conical micro centrifuge tube for 4 hours at 4° C.Protein A/G-coupled agarose beads (40 μL, of a 50% suspension) are thenadded to the tube. Following overnight tumbling at 4° C., the beads arewashed 3 times with ice-cold phosphate buffered saline. The beads arethen prepared for SDS-PAGE with 40 μL of Laemmli sample buffer. Proteinsseparated on SDS-PAGE are transferred onto nitrocellulose sheets withXCell-II Blot Module system (Invitrogen, Carlsbad, Calif.). The blotsare blocked with 5% BSA prior to incubation with a rabbit antibody toHIF-1α at 1:100 dilution (Novus). The blots are then washed withTris-buffered saline/Tween-20 buffer and incubated with horseradishperoxidase-conjugated goat anti-rabbit secondary antibody (Pierce,Rockford, Ill.). Blots are developed with the ECL reagent (Amersham,Piscataway, N.J.). Images of blots are captured with an Epson Expression1600 scanner.

Table VIII below provides non-limiting examples of the in vivo responsefor compounds according to the present disclosure, for example, HIFPH2(EGLN1) inhibition and VEGF stimulation.

TABLE VIII HIFPH2 VEGF No. Compound IC₅₀ (μM) IC₅₀ (μM) C17

11 27.4 1-benzyl-3-hydroxy-4-[(4- methylcyclohexylamino)methyl]pyridin-2(1H)-one C35

12 42.5 1-benzyl-3-hydroxy-4-{[3-(1H-imidazol-1-yl)propylamino]methyl}pyridin-2(1H)- one C14

12 20.6 1-benzyl-3-hydroxy-4-[(1- phenylethylamino)methyl]pyridin-2(1H)-one B5

9 53 (R)-1-benzyl-3-hydroxy-4-{[2- (methoxymethyl)pyrrolidin-1-yl]methyl}pyridin-2(1H)-one C33

16 53 1-benzyl-3-hydroxy-4-{[2- (methylthio)ethylamino]methyl}pyridin-2(1H)-one B14

11 78 1-benzyl-3-hydroxy-4-{[4-(6- chloropyridazin-3-yl)piperazin-1-yl]methyllpyridin-2(1H)-one C19

12 62.9 3-[(1-benzyl-3-hydroxy-2-oxo-1,2- dihydropyridin-4-yl)methylamino]azepan-2-one B9

17 12.6 1-benzyl-3-hydroxy-4-[(4- benzylpiperazin-1-yl)methyl]pyridin-2(1H)-one A18

18 29.2 1-(2-methoxybenzyl)-3-hydroxy-4-(azepan-1-ylmethyl)pyridin-2(1H)-one D10

4.4 27 1-[(4-methylphenyl)sulfonyl]-3-hydroxy-4-(1,4′-bipiperidin-1′-ylmethyl)pyridin- 2(1H)-one D14

12 19 1-[(4-methylphenyl)sulfonyl]-3-hydroxy-4-{[4-(6-chloropyridazin-3-yl)piperazin- 1-yl]methyl}pyridin-2(1H)-oneC1

12 42 1-(4-chlorobenzyl)-3-hydroxy-4-[(4-benzylamino)methyl]pyridin-2(1H)-one A41

14 16.6 tert-butyl 4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4- yl]methyllpiperazine-1-carboxylateE33

21 2.1 2-{[(3-hydroxy-2-oxo-1-tosyl-1,2- dihydropyridin-4-yl)methyl](methyl)amino}acetic acid F3

1.2 7.4 1-(4-chlorobenzyl)-3-hydroxypyridin- 2(1H)-one F2

5 >100 1-(3-chlorobenzyl)-3-hydroxypyridin- 2(1H)-one

Compound F2 was further tested in the mouse serum EPO assay describedherein above and found to have an EPO EC₅₀=14 μM.

Enhanced Neutrophil Activity

One aspect of the disclosure relates to the increased neutrophilactivity and increased neutrophil life that the disclosed compounds canprovide. The following provides methods and examples of increasedphagocytosis by the disclosed compounds. In the examples below theStaphylococcus aureus Newman cell strain is ATCC #25904 and themethicillin resistant Staphylococcus aureus strain is ATCC #33591, andthe U937 cell line is ATCC # CRL-1593.2. The HaCaT cells were generatedby the procedure of Boukamp P et al., “Normal keratinization in aspontaneously immortalized aneuploid human keratinocyte cell line.” J.Cell Biol. (1988) Mar:106(3):761-71.

For bacterial assays, S. Aureus (ATCC 33591) can be grown in Todd-Hewittbroth (THB) to logarithmic phase (OD₆₀₀ of 0.4 or ˜5×10⁷ cfu/mL) andthen pelleted, washed, and resuspended in PBS or RPMI 1640tissue-culture medium to the desired concentration. Venous blood fromhealthy volunteers can be used for whole blood and neutrophil isolation.Neutrophils can be purified using the PolyMorphPrep Kit (Axis Shield) inaccordance with manufacturer's instructions. Human monocytic cell lineU937 can be propagated in RPMI 1640 plus 10% fetal calf serum, 1 mmol/LNaPyr, 10 mmol/L HEPES, and glucose. Whole blood or phagocytic cells canbe preincubated with mimosine (Sigma-Aldrich) (0-500 μmol/L) for 2-4hours then challenged with S. Aureus (either 10⁵ cfu in 100 μL added to300 μL of whole blood or at an MOI of 1 bacterium/cell for isolatedphagocytes). Aliquots are then plated on THB agar after 30 (whole bloodand neutrophils) or 60 (U937 monocytes) min for enumeration of survivingS. Aureus colony-forming units.

Example 5

Isolated human neutrophils were pre-incubated for 1 hour at 37° C. witha control consisting of dimethyl sulfoxide (DMSO), 50 μM and 200 μM of acompound disclosed in Table VIII. Staphylococcus aureus (Newman strain)was then added to the neutrophils at an MOI of approximately 0.1 (1bacterium for every 10 neutrophils). Samples were taken at 60 and 90minutes wherein the neutrophils were lysed with water, and the totalbacteria remaining were enumerated on Todd-Hewitt broth (THB) agarplates.

FIG. 2 depicts the effectiveness of a compound disclosed in Table VIIIin providing enhanced killing of S. aureus (Newman strain) atconcentrations of 50 μM and 200 μM versus control. As can be seen inFIG. 2, at 90 minutes post-infection, approximately half of the colonyforming units are absent at a concentration of 200 μM.

Example 6

Cells from the human monocyte cell line U937 were pre-incubated for 2hours at 37° C. under an atmosphere of 5% CO₂ with a control consistingof DMSO and 10 μM of a compound disclosed in Table VIII. Staphylococcusaureus (virulent Newman strain) was then added to the cells at an MOI ofapproximately 1 (1 bacterium for every 1 cell). Samples are drawn at 30,60, 90 and 120 minutes post-infection. The U937 cells were lysed withTriton™, and the amount of bacteria remaining were enumerated on THBagar plates.

As depicted in FIG. 3, 4-prolyl hydroxylase inhibitor a compounddisclosed in Table VIII is effective in killing S. aureus when comparedto a control (DMSO). At 120 minutes, a compound disclosed in Table VIIIproduces an 84% kill of Newman strain S. aureus when the monocyte cellsare treated with 10 μM of a compound disclosed in Table VIII, therebyshowing increased phagocytosis due to extended neutrophil life span.

Example 7

Two samples of cells from the human monocyte cell line U937 werepre-treated with 10 μM of a compound disclosed in Table VIII. One samplewas pre-incubated for 1 hour and the other sample pre-incubated for 2hours, both at 37° C. under an atmosphere of 5% CO₂ . S. aureus(virulent Newman strain) was then added to the cells at an MOI ofapproximately 1-2 (1-2 bacteria for every 1 cell). Aliquots of cellswere removed from each sample at 30, 60, 90, and 120 minutespost-infection, the U937 cells were immediately lysed with Triton™, andtotal of remaining bacteria remaining were enumerated on THB agarplates.

As depicted in FIG. 4, U937 monocyte cells pre-treated with 10 μM of acompound disclosed in Table VIII for 1 hour (black bars) had almost nocolony forming units present 120 minutes post-infection, whereas thecells pre-treated two hours prior to infection had approximately 15%colony forming units present as compared to cells that were untreated.In addition, FIG. 4 indicates that within 1 hour after the U937 monocytecells had bee exposed to S. aureus (Newman strain), the number of colonyforming units present was significantly reduced relative to cellsreceiving no HIF-1α inhibitor.

Example 8

Two samples of cells from the human monocyte cell line U937 werepre-treated with 10 μM of a compound disclosed in Table VIII for 1 hourat 37° C. under an atmosphere of 5% CO₂ . S. aureus (Newman strain) wasadded to one sample and to the other was added methicillin-resistant S.aureus (MRSA). Both bacteria were added at an MOI of approximately 2-3(2-3 bacteria for every 1 cell). Aliquots of cells were removed fromeach sample at 30, 60, 90, and 120 minutes post-infection. The U937cells were immediately lysed with Triton™, and total of remainingbacteria remaining were enumerated on THB agar plates.

As depicted in FIG. 5, at 120 minutes post-infection, the MRSA infectedcells had only 25% of the average percentage of colony forming unitspresent when compared to control as represented by the black bars. Alsodepicted in FIG. 5, at 60 minutes post-infection, the Newman strain ofS. aureus had only approximately 12% of the average percentage of colonyforming units present when compared to control, and almost no colonyforming units present at 120 minutes post-infection as represented bythe hatched bars.

Example 9

Two samples of cells from the human monocyte cell line U937 treated with10 μM a compound disclosed in Table VIII were infected with either S.aureus (Newman strain) and methicillin-resistant S. aureus (MRSA). Bothbacteria were added at an MOI of approximately 2-3 (2-3 bacteria forevery 1 cell). Aliquots of cells were removed from each sample at 30,60, 90, and 120 minutes post-infection. The U937 cells were immediatelylysed with Triton™, and total remaining bacteria were enumerated on THBagar plates.

As depicted in FIG. 6, even without pre-treatment with a compounddisclosed in Table VIII, at 60 minutes post-infection, the Newman strainof S. aureus had only 25% of the average percentage of colony formingunits present when compared to control as represented by the black bars.The MRSA strain was reduced to less than approximately 40% of theaverage percentage of colony forming units present when compared tocontrol as represented by the hatched bars.

Example 10

Three samples of cells from the human monocyte cell line U937 weretreated with 100 μM mimosine, 2 μg/mL vancomycin or 10 μM of a compounddisclosed in Table VIII. Each sample was infected with either S. aureus(Newman strain) or methicillin-resistant S. aureus (MRSA). Both bacteriawere added at an MOI of approximately 2-3 (2-3 bacteria for every 1cell). At 120 minutes post-infection aliquots were withdrawn from allsix samples and the U937 cells were immediately lysed with Triton™, andtotal remaining bacteria were enumerated on THB agar plates.

As depicted in FIG. 7, 10 μM a compound disclosed in Table VIII enhancedkill of both bacterial strains, i.e., S. aureus, Newman (hatched bars)or MRSA (black bars), when compared to mimosine treated cells. Referringto the hatched bars representing Newman strain, as further depicted inFIG. 7, the sample treated with 10 μM a compound disclosed in Table VIIIhad a lower average percentage of colony forming units present than thecells treated with vancomycin. The U937 cells infected with MRSA (blackbars) had approximately 40% of the colony forming units present versusuntreated cells and less than half the number of those treated withmimosine.

FIG. 8 depicts the average percentage of colony forming units present(Newman strain) versus control for human monocyte cells (U937) at 30,60, 90, and 120 minutes post-infection, when treated with 10 μM acompound disclosed in Table VIII. The black bars represent treatmentwith a compound disclosed in Table VIII beginning at the time ofinfection with S. aureus, the hatched bars represent cells pretreatedwith a compound disclosed in Table VIII and white bars represent cellspretreated two hours prior to infection with S. aureus.

FIG. 9 depicts the average percent of colony forming units present at120 minutes post-infection vs DMSO (control) when HaCaT cells arepre-treated for 1 hour according to the examples above with 800 μMmimosine, 10 μM of compound disclosed in Table VIII or 1 μg mLvancomycin followed by inoculation with S. aureus (Newman strain,hatched bars) and methicillin-resistant S. aureus (MRSA, black bars).FIG. 10 depicts the average percent of colony forming units present at30, 60, 90, and 120 minutes post-infection for Newman strain of S.aureus (hatched bars) and MRSA (black bars) when HaCaT cells arepre-treated for 1 hour according to the examples above with 10 μM of acompound disclosed in Table VIII.

FIG. 11 depicts the up regulation of phosphoglycerate kinase (PGK)expression in wild type murine embryonic fibroblasts as a result oftreatment with a compound disclosed in Table VIII at dosages of 1 μM(E), 10 μM (F), and 50 μM (G) vs. wild type control (H) and the lack ofup regulation of PGK expression in HIF-1 knock out cells as a result oftreatment with a compound disclosed in Table VIII at dosages of 1 μM(A), 10 μM (B), and 50 μM (C) and HIF-1 knock out control (D). Both celltypes were treated for 7 hours.

FIG. 12 depicts the up regulation of phosphoglycerate kinase (PGK)expression in wild type murine embryonic fibroblasts as a result oftreatment with a compound disclosed in Table VIII at dosages of 1 μM(E), 10 μM (F), vs. wild type control (G) and the lack of up regulationof PGK expression in HIF-1 knock out cells as a result of treatment witha compound disclosed in Table VIII at dosages of 1 μM (A), 10 μM (B),and 50 μM (C) and HIF-1 knock out control (D).

FIG. 13 depicts the up regulation of phosphoglycerate kinase (PGK)expression in wild type murine embryonic fibroblasts as a result oftreatment with a compound disclosed in Table VIII at dosages of 1 μM(E), 10 μM (F), and 50 μM (G) vs. wild type control (H) and the lack ofup regulation of PGK expression in HIF-1 knock out cells as a result oftreatment with a compound disclosed in Table VIII at dosages of 1 μM(A), 10 μM (B), and 50 μM (C) and HIF-1 knock out control (D).

Vascular Endothelial Growth Factor (VEGF) is dependent upon the presenceof HIF-1 in cells. FIG. 14 depicts the up regulation of vascularendothelia growth factor (VEGF) expression in wild type murine embryonicfibroblasts as a result of treatment with a compound disclosed in TableVIII at dosages of 1 μM (E), 10 μM (F), and 50 μM (G) vs. control (H)and the lack of up regulation of VEGF expression in HIF-1 knock outcells treated with a compound disclosed in Table VIII at dosages of 1 μM(A), 10 μM (B), and 50 μM (C) and HIF-1 knock out control (D). Both celltypes were treated for 7 hours. As seen in FIG. 14, VEGF is increasedwhen dosed at 10 μM (F) and 50 μM (G). In HIF-1 knock out cells, thereis no increase in PGK up regulation when HIF-1 knock out cells are dosedat 1 μM (A), 10 μM (B), and 50 μM (C) when compared to wild type control(H) and HIF-1 knock out control (D).

Wound Healing Example 11

Twenty-four (24) mice were divided into three groups. Group 2 animalswere administered bacterial inoculum (Staphylococcus aureus antibioticsensitive Newman strain [ATCC #25904]) by subcutaneous injection on Day0 and received 10 μM of a compound disclosed in Table VIII for 6 daysstarting at 2 hours post-infection (Days 0-5). Group 1 receivedsubcutaneous injections of DMSO. Group 3 served as a control group andreceived no treatment. Lesion size was monitored daily during the study.Only open wounds were considered lesions; bumps and white patcheswithout an open wound were not measured for lesion size. On Day 7, thefinal lesion size was measured and mice were sacrificed fordetermination of bacterial load in skin and kidney. Day 7post-infection, mice were sacrificed after final lesion size measurementand the lesioned skin tissue and both kidneys were collected. Skin andkidneys were homogenized in phosphate buffered saline, serially diluted,and plated on Todd-Hewitt agar plates to enumerate colony forming unitsof bacteria.

FIG. 15 shows the significant reduction in the size of skin lesions(wounds) for animals in Group 1 (solid circles (●)) treated with 10 μMof a compound disclosed in Table VIII versus animal treated with DMSO(solid squares (▪)). As depicted in FIG. 15, mice infected with Newmanstrain of S. aureus followed by treatment with 10 μM of a compounddisclosed in Table VIII or DMSO (control) at 2 hours post-infection. Thedata show the statistically significant reduction in the size of skinlesions (wounds) for animals treated with a compound disclosed in TableVIII (solid circles (●)) or DMSO (solid squares (▪)).

FIG. 16 shows the significant reduction in the size of skin lesions(wounds) for animals in Group 1 (solid circles (●)) treated with 10 μMof a compound disclosed in Table VIII versus untreated animals (solidtriangles (A)). As depicted in FIG. 16, mice infected with Newman strainof S. aureus followed by treatment with 10 μM of a compound disclosed inTable VIII or no treatment at 2 hours post-infection. The data show thereduction in the size of skin lesions (wounds) for animals treated witha compound disclosed in Table VIII (solid circles (●)) or untreated(solid triangles (▴)).

Example 12

Twenty-four (24) mice were divided into three groups. Group 1 animalswere administered bacterial inoculum (Staphylococcus aureus antibioticsensitive Newman strain [ATCC #25904]) by subcutaneous injection on Day0 and received 10 μM of a compound disclosed in Table VIII for 6 daysstarting at 2 hours post-infection (Days 0-5). Group 2 receivedsubcutaneous injections of DMSO. Group 3 served as a control group andreceived no treatment. Lesion size was monitored daily during the study.Only open wounds were considered lesions; bumps and white patcheswithout an open wound were not measured for lesion size. Day 7post-infection, mice were sacrificed after final lesion size measurementand lesioned skin tissue and both kidneys were collected. Skin andkidneys were homogenized in phosphate buffered saline, serially diluted,and plated on Todd-Hewitt agar plates to enumerate colony forming unitsof bacteria.

FIG. 17 is a plot histogram wherein the number of observed colonyforming units per gram of skin tissue is depicted. The straight linesindicate the mean value for each group. The results for the untreatedgroup are plotted under (A), the results for the group treated with DMSOare plotted under (B) and results for the group treated with 10 μM of acompound disclosed in Table VIII are plotted under (C).

FIG. 18 is a plot of the observed colony forming units of bacteria foundin the kidneys of the animals. The results for the untreated group areplotted under (A), the results for the group treated with DMSO areplotted under (B) and results for the group treated with 10 μM of acompound disclosed in Table VIII are plotted under (C). As can be seenfrom these data, half of the animals treated with the HIF-1α prolylhydroxylase inhibitor disclosed in Table VIII had no bacteria in thekidney indicating that the compound disclosed in Table VIII was able tosystemically prevent spread of the infection from the wound to thekidney.

Example 13

Twenty (20) mice were divided into two groups. Group 1 animals wereadministered bacterial inoculum (Streptococcus pyogenes NZ131 [M49strain]) by subcutaneous injection on Day 0 and were pretreated with acompound disclosed in Table VIII once per day for 4 days, starting 2hours pre-infection (Days 0-3). A compound disclosed in Table VIII wasformulated in cyclodextran and diluted in distilled water prior tosubcutaneous injection, at a dose of 0.5 mg/kg. Lesion size wasmonitored daily during the study. Only open wounds were consideredlesions; bumps and white patches without an open wound were not measuredfor lesion size. On Day 4 post-infection, mice were sacrificed afterfinal lesion size measurement and lesioned skin tissue and both kidneyswere collected. Skin and kidneys were homogenized in phosphate bufferedsaline, serially diluted, and plated on Todd-Hewitt agar plates toenumerate colony forming units of bacteria.

FIG. 19 depicts the results of Example 13 wherein 2 groups of animalsare treated with Streptococcus pyogenes NZ131 [M49 strain]. The datashow the reduction in the size of skin lesions (wounds) for animals inGroup 1 (solid triangles (▴)) treated with 0.5 mg/kg of a compounddisclosed in Table VIII versus animal treated with vehicle control(cyclodextran) (solid circles (●)). FIG. 20 is a plot histogram thatalso depicts the results of Example 12 wherein the number of colonyforming units for the observed skin lesions on animals treated withvehicle control (cyclodextran) are plotted under (A) and results for thegroup treated with 0.5 mg/kg of a compound disclosed in Table VIII areplotted under (B).

Kits

Also disclosed are kits comprising the HIF-1α prolyl hydroxylaseinhibitors be delivered into a human, mammal, or cell. The kits cancomprise one or more packaged unit doses of a composition comprising oneor more HIF-1α prolyl hydroxylase inhibitors to be delivered into ahuman, mammal, or cell. The units dosage ampules or multidosecontainers, in which the HIF-1α prolyl hydroxylase inhibitors to bedelivered are packaged prior to use, can comprise an hermetically sealedcontainer enclosing an amount of polynucleotide or solution containing asubstance suitable for a pharmaceutically effective dose thereof, ormultiples of an effective dose. The HIF-1α prolyl hydroxylase inhibitorcan be packaged as a sterile formulation, and the hermetically sealedcontainer is designed to preserve sterility of the formulation untiluse.

The disclosed HIF-1α prolyl hydroxylase inhibitors can also be presentin liquids, emulsions, or suspensions for delivery of active therapeuticagents in aerosol form to cavities of the body such as the nose, throat,or bronchial passages. The ratio of HIF-1αprolyl hydroxylase inhibitorsto the other compounding agents in these preparations will vary as thedosage form requires.

Depending on the intended mode of administration, the pharmaceuticalcompositions can be in the form of solid, semi-solid or liquid dosageforms, such as, for example, tablets, suppositories, pills, capsules,powders, liquids, suspensions, lotions, creams, gels, or the like,preferably in unit dosage form suitable for single administration of aprecise dosage. The compositions will include, as noted above, aneffective amount of the HIF-1α prolyl hydroxylase inhibitor incombination with a pharmaceutically acceptable carrier and, in addition,can include other medicinal agents, pharmaceutical agents, carriers,adjuvants, diluents, etc.

For solid compositions, conventional nontoxic solid carriers include,for example, pharmaceutical grades of mannitol, lactose, starch,magnesium stearate, sodium saccharin, talc, cellulose, glucose, sucrose,magnesium carbonate, and the like. Liquid pharmaceutically administrablecompositions can, for example, be prepared by dissolving, dispersing,etc., an active compound as described herein and optional pharmaceuticaladjuvants in an excipient, such as, for example, water, saline aqueousdextrose, glycerol, ethanol, and the like, to thereby form a solution orsuspension. If desired, the pharmaceutical composition to beadministered can also contain minor amounts of nontoxic auxiliarysubstances such as wetting or emulsifying agents, pH buffering agentsand the like, for example, sodium acetate, sorbitan monolaurate,triethanolamine sodium acetate, triethanolamine oleate, etc. Actualmethods of preparing such dosage forms are known, or will be apparent,to those skilled in this art; for example see Remington's PharmaceuticalSciences, referenced above.

Parenteral administration, if used, is generally characterized byinjection. Injectables can be prepared in conventional forms, either asliquid solutions or suspensions, solid forms suitable for solution orsuspension in liquid prior to injection, or as emulsions. A morerecently revised approach for parenteral administration involves use ofa slow release or sustained release system, such that a constant levelof dosage is maintained. See, e.g., U.S. Pat. No. 3,710,795, which isincorporated by reference herein.

When the HIF-1α prolyl hydroxylase inhibitors are to be delivered into amammal other than a human, the mammal can be a non-human primate, horse,pig, rabbit, dog, sheep, goat, cow, cat, guinea pig or rodent. The termshuman and mammal do not denote a particular age or sex. Thus, adult andnewborn subjects, as well as fetuses, whether male or female, areintended to be covered. A patient, subject, human or mammal refers to asubject afflicted with a disease or disorder. The term “patient”includes human and veterinary subjects.

While particular embodiments of the present disclosure have beenillustrated and described, it would be obvious to those skilled in theart that various other changes and modifications can be made withoutdeparting from the spirit and scope of the disclosure. It is thereforeintended to cover in the appended claims all such changes andmodifications that are within the scope of this disclosure.

What is claimed is:
 1. A method for treating a wound in a subject,comprising administering to a subject an effective amount of a compoundhaving the formula:

wherein Z is phenyl substituted with from 1 to 5 halogens chosen fromfluorine and chlorine; R⁴ is C₁-C₄ linear alkyl or C₃-C₄ branched alkyl;or a pharmaceutically acceptable salt thereof.
 2. The method accordingto claim 1, wherein R⁴ is methyl.
 3. The method according to claim 1,wherein R⁴ is ethyl.
 4. The method according to claim 1, wherein R⁴ istert-butyl.
 5. The method according to claim 1, wherein Z is4-chlorophenyl.
 6. The method according to claim 1, wherein Z is chosenfrom 2-chlorophenyl, 3-chlorophenyl, 2-fluorophenyl, 3-fluorophenyl, or4-fluorophenyl.
 7. The method according to claim 1, wherein Z is chosenfrom 2,3-difluorophenyl, 2,4-difluorophenyl, 2,5-difluorophenyl,2,6-difluorophenyl, 2,3-dichlorophenyl, 2,4-dichlorophenyl,2,5-dichlorophenyl, and 2,6-dichlorophenyl.
 8. The method according toclaim 1, wherein the compound is tert-butyl4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate.9. The method according to claim 1, wherein the compound chosen from:Methyl4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(3-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(2-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(4-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(3-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(2-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;tert-Butyl4-{[1-(3-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;tert-Butyl4-{[1-(2-chlorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(4-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(3-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(2-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(4-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(3-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(2-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;tert-Butyl4-{[1-(4-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;tert-Butyl4-{[1-(3-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;tert-Butyl4-{[1-(2-fluorobenzyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(4-chlorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(3-chlorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(2-chlorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(4-chlorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(3-chlorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(2-chlorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;tert-Butyl4-{[1-(3-chlorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;tert-Butyl4-{[1-(2-chlorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(4-fluorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(3-fluorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Methyl4-{[1-(2-fluorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(4-fluorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(3-fluorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;Ethyl4-{[1-(2-fluorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;tert-Butyl4-{[1-(4-fluorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;tert-Butyl4-{[1-(3-fluorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate;and tert-Butyl4-{[1-(2-fluorophenylsulfonyl)-3-hydroxy-2-oxo-1,2-dihydropyridin-4-yl]methyl}piperazine-1-carboxylate.10. The method according to claim 1, wherein the compound is apharmaceutically acceptable salt of an anion chosen from chloride,bromide, iodide, sulfate, bisulfate, carbonate, bicarbonate, phosphate,hydrogensulfonate, p-toluenesulfonate, methanesulfonate, formate,acetate, propionate, butyrate, pyruvate, lactate, oxalate, malonate,maleate, succinate, tartrate, fumarate, glycolate, or citrate.